Bioprocessing Residue Detection Solutions

www. aero bi osystem s. com Scan the QR code to download this resource Biopharmaceutical process residue detection is a quality control methodology used to quantify the number of residues generated throughout the biological manufacturing process. The production of biologics such as cell and gene therapies, therapeutic antibodies, and vaccines involves the use of biological materials. As such, residues with a biological origin such as host cell DNA, affinity chromatography ligands, culture medium additives, and other substances may be introduced. It is crucial to remove and quantitatively control these introduced residues during the production process for drug/vaccine manufacturing, as residual contaminants can severely impact the safety and efficacy of the final product. resDetect™ is our brand of reagent kits designed to follow international legal and regulatory requirements and guidelines for the detection of residual components. These kits cover various applications related to process-related residue detection for biopharmaceuticals, including antibody drugs, cell and gene therapies (CGT), vaccines, and more. resDetect™ offers comprehensive solutions for process residue detection and includes reagent kits for the detection of residual components such as host cell DNA, host cell protein, host cell RNA, protein A, tool enzymes, cytokines, and more. \?.) 􀀂, Stable and Reliable -- Stringent batch testing and release, with inter-batch differences less than 15%. Regulatory Compliance -- Complaitn with standards and regulations of international regulatory agencies. Global Supply -- Product delivery in 1-3 days, worldwide. Comprehensive Validation -- Referencing guidelines including ICH Q2 (R2) and ChP 9101. Cell therapy process residue quality control Gene therapy process residue quality control Therapeutic antibodies process residue quality control Tel: +l 800-810-0816 • Web: www.acrobiosystems.com mRNA-based therapeutics process residue quality control Email: order@acrobiosystems.com ' • Tel: +l 800-810-0816 • Web: www.acrobiosystems.com Biopharmaceutical CMC {Chemistry, Manufacturing, and Controls) residue quality control Cell therapy process residue quality control Host cell nucleic acid residue detection Plasmid DNA residue detection Viral oncogenes residue detection Cytokine residue detection Antibiotic residue detection Enzyme residue detection Gene therapy process residue quality control Host cell nucleic acid residue detection Plasmid DNA residue detection Viral oncogenes residue detection Nuclease residue detection Antibiotic residue detection Therapeutic antibodies process residue quality control Affinity ligand residue detection Host cell nucleic acid residue detection mRNA-based therapeutics process residue quality control Host cell residual nucleic acid detection Enzyme residue detection dsRNA residue detection Email: order@acrobiosystems.com p04 Nucleic Acid Residue Host Cell Residual Nucleic Acid Detection Types of Residues in CMC Process Quality Control Plasmid DNA Residue Detection Viral Oncogenes Residue Detection p09 Affinity Ligand Residue Protein A Residue Detection p12 Process Additive Residue Cytokine Residue Detection Antibiotic Residue Detection Enzyme Residue Detection p18 Other Residues DNAse Residue Detection RNAse Residue Detection Tel: +l 800-810-0816 • Web: www.acrobiosystems.com Email: order@acrobiosystems.com [ Automatic Nucleic Acid Extraction System -- - • 􀀶 Purify up to 32 samples within 40 minutes. 􀀶 Equipped with door opening protection and UV disinfection measures to prevent contamination. 􀀶 Permanent magnetic bars with dynamically adjustable amplitude, ensuring residue-free and wall-free extraction. 􀀶 Precision automation operation, high extraction efficiency, stable results, and small inter-hole difference. 􀀶 Compatible with various magnetic bead-based nucleic acid extraction reagent kits. 􀀶 3Q certification, audit tracking version (customizable). The fully automated nucleic acid pretreatment system is a magnetic bead-based automated DNA sample processing system. Paired with our sample preparation reagent kits, it can reliably, efficiently, and automatically extract residual DNA from samples. [ resDe;ect™ Host Cell Residual DNA Detection Kit 􀀶 Host cell DNA types: HEK293, E. coli, CHO, etc. 􀀶 High sensitivity: Developed based on quantitative PCR methods, with a Limit of Detection (LOD) reaching 1 fg/μL. 􀀶 High consistency: Consistent performance across different DNA fragment sizes. 􀀶 High specificity: No cross-reactivity with unrelated DNA, minimizing false positive detection. 􀀶 Manufactured under GMP-like facility and strictly adheres to an ISO13485:2016, ISO9001 :2015 quality management system. 􀀶 Rigorous quality management system; comprehensive validation: References to ICH Q2(R2) guidelines with thorough validation reports. ■ Product List Cat. No. OPE-32S OPA-R00S OPA-0002 OPA-R004 OPA-R006 OPA-R010 OPA-R009 ■ Product Data Pro duct Description resDetect'" Automated Nucleic Acid Extraction System resDetect'" resDNA Sample Preparation Kit (Magnetic Beads) resDetect'" E coli resDNA Quantitative Kit (qPCR) resDetect'" CHO resDNA Quantitation Kit (qPCR) resDetect'" HEK293 resDNA Quantitation Kit (qPCR) resDetect'" HEK293T resDNA Quantitation Kit (qPCR) resDetect'" Plasmid resDNA Quantitation Kit (qPCR) ► High Sensitivity: Limit of Detection (LLOD) reaches 1 fg/μL A .. I" Standard 1 Standard 2 Standard 3 Standard4 Standard 5 St.::mdard 6 B R2 = 0.999 Eff% = 99.851 High sensitivity and broad dynamic range using the E.coli resDNA Quantitative Kit (qPCR) (Cat. No. OPA-O002). (A) Typical analysis results obtained with Standard 1 (300 pg/μL) to 6 (3 fg/μL). (B) The standard curve of the 10-fold dilution series. PCR efficiency should be 90-110%. Tel: +l 800-810-0816 • Web: www.acrobiosystems.com Email: order@acrobiosystems.com ► High Specificity: No cross-reactivity with unrelated DNA A ■ Eco/I ■ Ecol,..-CHO R2 = 0.999 Eff% = 93.255 B : ■ Ecolt ■ £􀀢HEK293 ' ' . " . . . . - R2 = 0.999 Eff% = 91.125 Assay specificity. Standard curves generated using 1 0-fold serial dilution of E. coli genomic DNA (included in the kit). ► High Consistency: Maintains consistent performance across different DNA fragment sizes gDNA R' = 0.999 Effli = 99.741 Smin R' = 0.999 Effli =104.106 13min 20 min iL_􀀅 . . . . . . . . . . . . . . . . . . R' = 0.999 ,... R' = 0.999 Effli =105.965 •--......_ Effli =102.462 Consistent quantitation across a broad range of fragment sizes. Standard curves were generated using a 10-fold serial dilution of gDNA and fragmented DNA. Fragmented DNA was generated by sonicating total E. coli genomic DNA (8min, 1 3min, 20min). Fragmentation of the DNA was confirmed by agarose gel analysis. ► Competitive Comparison Data :r-. Compe,;,o,H 1 Amplification curves Standard curves __ l?@ __ ·L - - L0· R' = 0.999 Efflo = 99.851 R1 : 0.999 Elfli= 88.795 The amplification efficiency of standard curve (ACRO) is close to 100%, while the amplification efficiency of standard curve (Competitor H) is less than 90%. Better performances can be observed based on the standard curves from each kit. [ resDetect™ Viral Oncogenes Residual DNA Detection Kit Designed specifically to quantify residual E1A and SV40LTA DNA in biopharmaceutical production (cells, viruses, etc.), this kit is based on a real-time quantitative PCR (qPCR) method. It can rapidly and reliably detect the content of residual DNA, with a quantitative lower limit of 40 copies/μL. Typically, all operations can be completed within 4 hours. It can be used in conjunction with the resDNA Sample Preparation Kit (Magnetic Bead Method) (Catalog Number: OPA-R00S) for DNA extraction to achieve optimal detection performance. ■ Product list res Detect™ E1 A res DNA Quantitation Kit (qPCR) OPA-R007 res Detect™ E1 A & SV40L TA res DNA Quantitation Kit (qPCR) Tel: +l 800-810-0816 • Web: www.acrobiosystems.com Email: order@acrobiosystems.com ■ Product Data ► High Sensitivity A R2: 0.999 Eff%: 98.65 Slope: -3.355 High sensitivity and broad dynamic range using the E1A&SV40L TA resDNA Quantitative Kit. (A) Typical analysis results of obtained with E1A DNA Control Standard 1 (4x1Q• copies/μL) to 6 (4x10 copies/μL). (B) A standard curve of the E1A DNA Control 10-fold dilution series. PCR efficiency should be within 90-110%. (C) Typical analysis results of obtained with SV40L TA DNA Control Standard 1 (4x 1 o• copies/μL) to 6 (4x 10 copies/μL). (D) The standard curve of the SV40L TA DNA control 10-fold dilution series. PCR efficiency should be 90-110%. ► High Specificity Unrelated DNA E coli DNA CHO DNA Vero DNA Pichia pastoris DNA MDCKDNA Mean (copies/μL) E1A 0.99 Undetermined Undetermined Undetermined 1.76 SV40LTA 1.52 Undetermined Undetermined Undetermined 1.26 To evaluate the specificity using E1A&SV40L TA resDNA Quantitation Kit, samples spiked with unrelated DNA were tested. Results shown in the following Table. Unrelated DNA (CHO, Vero, Pichia pastoris) were not detected in assay, and the values of E. coli and MOCK were outside the range of standard curve (4x 106 copies/μL to 4x 10 copies/μL). Samples (copies/μL) Mean (copies/μL) Recovery (%) E1A SV4OLTA E1A SV4OLTA 4x106 3.94x106 3.64x106 98.4 90.93 4x104 4.00x104 3.40x104 99.89 84.98 4x10 3.52x10 3.14x10 87.97 78.57 To evaluate the recovery of assay using E1A&SV40L TA resDNA Quantitation Kit, three different concentrations of DNA samples were tested. Results shown in the following Table. All samples had recoveries between 78%-100%. [ resDetect™ Plasmid DNA Residual Detection Kit ACROBiosystems' resDetect™ Plasmid DNA Residual Detection Kit (Catalog Number: OPA-R009) enables quantitative detection of residual plasmid DNA in various bioproducts, including intermediates, semi-finished products, and finished products. Tel: +l 800-810-0816 • Web: www.acrobiosystems.com Email: order@acrobiosystems.com ■ Product Data ► High Sensitivity Linearize DNA control Standardl Standard2 Standard3 Standard4 Standards Standard6 R': 0.999 Effll: 104.33 Slope: -3.222 R2: 0.999 Elf%: 106.129 Slope: -3.183 High sensitivity and broad dynamic range using the Plasmid resDNA Quantitation Kit. (A) Typical analysis results of obtained with using linearized DNA control Standard 1 (4x1Q• copies/μL) to 6 (4x10 copies/μL). (B) The standard curve of the Linearize DNA control 10-fold dilution series. PCR efficiency should be 90-110%. (C) Typical analysis results of obtained with circular DNA control Standard 1 (2x 1 06 copies/μL) to 5 (2x 102 copies/μL). (D) The standard curve of the Circular DNA control 1 0-fold dilution series. PCR efficiency should be 90- 1 1 0%. ► High Specificity Unrelated DNA HEK293 DNA CHO DNA Vero DNA Pichia pastoris DNA MDCK DNA Mean (copies/μL) Plasmid 5.08 0.88 0.99 2.8 23.86 To evaluate the specificity of assay using Plasmid resDNA Quantitation Kit, no template samples spiked with unrelated DNA were tested. Results shown in the following Table. Unrelated DNA (HEK293, CHO, Vero, Pichia pastoris, and MOCK) were detected in assay, however the values of Mean were outside the range of standard curve (4x 1 06 copies/μL to 4x 1 0 copies/μL). Linearize Samples (copies/μL) Mean (copies/μL) Recovery (%) Plasmid Plasmid 4x106 4.34x106 108.61 4x104 4.33x104 108.22 4x10 3.76x10 94.07 Circular Samples (copies/μL) Mean (copies/μL) Recovery (%) Plasmid Plasmid 2x106 1.88x106 93.86 2x104 1.71x104 85.40 2x102 1.87x102 93.35 To evaluate the recovery of assay using Plasmid resDNA Quantitation Kit, three different concentrations of DNA samples were tested. Results shown in the following Table. All samples had recoveries between 80%-110%. Tel: +l 800-810-0816 • Web: www.acrobiosystems.com Email: order@acrobiosystems.com [ resDetect™ Universal Protein A Quick ELISA kit ACROBiosystems' resDetect™ Universal Protein A Quick ELISA kit (Catalog Number: RES-A024) was developed to support drug CMC production and quality control-related research. Through rigorous methodological validation, this kit is based on sandwich enzyme-linked immunosorbent assay (sandwich ELISA). It can be used to determine the presence of native or structurally conserved recombinant Protein A in intermediate as well as final products of antibodies and Fe fusion protein drug formulations. It also detects the dissociation of various alkali-resistant recombinant Protein A ligands commonly used in bioprocess manufacturing applications, such as MabSelect SuRe™ and MaXtar® ARPA Sandwich ELISA ligand. The kit meets the needs of pharmaceutical companies for residual quantitative analysis and optimization of relevant drug purification processes, thereby expediting the process of bringing biopharmaceuticals to market. ■ Application The kit is developed for the detection of natural or structurally conserved recombinant forms of Protein A and alkaline-resistant Protein A variants, such as MabSelect SuRe™ , MaXtar® ARPA ligand (Bio-Link Co.), etc. in bioprocess manufacturing applications. ■ Features g" Universal - Suitable for detection of natural or structurally conserved recombinant forms of Protein A and alkaline-resistant Protein A variants, such as MabSelect Su Re TM and other ligands g" Fast time to results - less than 2 hours g-Accuracy - Tracebility of Protein A standards against BSA China National Standard (NIFDC code: 140619) with validated pharmacopoeia quantitation methodology g" Extensive validation - Validation Report (ICH compliant) available on request g" High sensitivity - Sensitivity < 20 pg/ml of recombinant Protein A and MabSelect™ SuRe Protein A or other Protein A ligands g" High lgG tolerance - Accurately quantify protein A in up to 10 mg/ml antibody g" Excellent buffer compatibility ■ Product Data ► Applicability This kit includes three different types of Protein A standards: Recombinant Protein A Standard, Alkali-Tolerant Recombinant Protein A Standard, and MaXtar® ARPA ligand Protein A Standard (Bio-Link Co.). For the detection of ligand residues in different Protein A affinity chromatography resins, the recommended standards are as follows: Resin MabSelect Su Re™ MaXtar® ARPA ligand AlkaliT- olerant Protein A Resin 1 (Manufacturer B) AlkaliT- olerant Protein A Resin 2 (Manufacturer D) AlkaliT- olerant Protein A Resin 3 (Manufacturer N) native & rproteIn A Standards Alkali-Tolerant Recombinant Protein A Standard MaXtar® ARPA ligand Protein A Standard (Bio-Link Co.) Alkali-Tolerant Recombinant Protein A Standard Alkali-Tolerant Recombinant Protein A Standard Alkali-Tolerant Recombinant Protein A Standard Recombinant Protein A Standard Tel: +l 800-810-0816 • Web: www.acrobiosystems.com Email: order@acrobiosystems.com * ACROBiosystems' Protein A Residue Detection Kit has excellent recovery rates (80-1 20%). In addition to MabSelect SuRe™ and MaXtar® ARPA ligand, three alkali-tolerant affinity media commonly used in the market were also tested. The data shows ideal recovery rates and compatibility, demonstrating excellent versatility. Ligand Competitor Natural and Recom binant Protein A M a bSelect Su Re™ Protein A MaXtar' ARPA ligand Protei n A (Bio-Link Co.) Alkali-Tolerant Protei n A Resin 1 ( M anufacturer B) Alkali-Tolera nt Protei n A Resin 2 ( M a nufacturer D) Alkali-Tolera nt Protei n A Resin 3 ( M a nufacturer N) ► Sample Data ACR0 93-106% 89-106% 91-112% 90-101% 96-105% 96-121% Competitor C Competitor P 125-329% 68-103% 106-148% 105-158% 146-186% 93-127% 176-213% 131-170% 83-118% 57-121% 122-164% 96-154% * Recognition and detection of various Protein A structures, including native or structurally conserved recombinant Protein A, as well as various alkali-resistant Protein A ligands commonly used in bioprocess manufacturing applications, such as MabSelect SuRe™, MaXtar® ARPA ligand, etc. St a n d a rd N u m . C o n c e ntrat i o n Standard 7 1 . 6 ng/ml Standard 6 0.8 ng/ml Standard 5 0.4 ng/ml Standard 4 0.2 ng/ml Standard 3 0.1 ng/ml Standard 2 0.05 ng/ml Standard 1 0.025 ng/ml Standard 0 0 ng/ml O D4 s o n m 3.032 3 ----- 2. 01 7 1 . 1 44 0.624 0.342 0.205 0. 1 43 0.076 --- -- -- -----·-'·-·- ··· -·- - ----'- R2,,P.99993 , 1.5 Protein A Cone. (ng/.t..) The standard curve is generated by diluting the AGRO self-produced high-purity, high-sensitivity recombinant Protein A series and fitting the data using a 4-parameter logistic (4-Pl) model. The detection range is from 25 to 1 600 pg/ml, with an R2 value of up to 0.99. St a n d a rd N u m . C o n c e ntrat i o n Standard 7 1 . 6 ng/ml Standard 6 0.8 ng/ml Standard 5 0.4 ng/ml Standard 4 0.2 ng/ml Standard 3 0.1 ng/ml Standard 2 0.05 ng/ml Standard 1 0.025 ng/ml Standard 0 0 ng/ml 0 0 450nm 3. 1 97 2.41 5 1 .4 0.795 0.474 0.267 0. 1 82 0.092 ' . 1 1.5 P,otein A Cone. (nghnll The standard curve is generated by diluting the AGRO self-produced high-purity, high-sensitivity alkali-tolerant recombinant Protein A series and fitting the data using a 4-parameter logistic (4-Pl) model. The detection range is from 25 to 1 600 pg/ml, with an R' value of up to 0.99. St a n d a rd N u m . C o n ce ntrati o n Standard 7 1 .6 ng/ml Standard 6 0.8 ng/ml Standard 5 0.4 ng/ml Standard 4 0.2 ng/ml Standard 3 0.1 ng/ml Standard 2 0.05 ng/ml Standard 1 0.025 ng/ml Standard 0 0 ng/ml 0 0 450nm 2.945 2.052 1 . 1 94 0.635 0.358 0.21 2 0. 1 4 0.068 2.5 ..... 0.5 1 1.5 P1otein A Cone. (ng/ml) The standard curve is generated by diluting the high-purity, high-sensitivity MaXtar" ARPA ligand Protein A series self-produced by Bio-Link, and fitting the data using a 4-parameter logistic (4-Pl) model. The detection range is from 25 to 1600 pg/ml, with an R' value of up to 0.99. Tel: +l 800-810-0816 • Web: www.acrobiosystems.com Email: order@acrobiosystems.com [ resDetect™ Cytokine/Antibody Residue Detection Kit In the production process of immune cell therapy products, culture medium supplements such as GMP-grade IL-2, IL-15, IL-7, and IL-21, among others, are essential reagents for the proliferation and differentiation of immune cells such as T/NK cells. They are key raw materials for the production of immune cell therapy drugs. However, to ensure the safety of the final product, it is necessary to control the residual amounts of these culture additives. ■ Hot Products list Cat. No. Product Description - , CRS-A024 CRS-A025 CRS-A003 CRS-A005 CRS-A01 0 CRS-A01 4 CRS-A01 5 resDetect™ Human lnterleukin-1 5 (IL-1 5) ELISA Kit (Residue Testing) resDetect™ Human lnterleukin-7 (IL-7) ELISA Kit (Residue Testing) resDetect™ Human lnterleukin-2 (IL-2) ELISA Kit (Residue Testing) resDetect™ Human lnterleukin-6 (IL-6) ELISA Kit (Residue Testing) resDetect™ Human lnterleukin-21 (IL-21 ) ELISA Kit (Residue Testing) resDetect™ Anti-CD28 Antibody ELISA Kit Learn more i nformation resDetect™ Anti-CD3 Antibody ELISA Kit ■ Product Data ► Sample Data R2=0-2998 -/'---,----------, For each experiment. a standard curve needs to be set for each micro-plate. and the specific OD value may vary depending on different laboratories. testers, or equipments. The following example data is for reference only (CRS-A024) . Conc.(po/ml) ► Dilution Linearity Cell culture medium Average Recovery (%) 1 02.6 1 :2 Range (%) 97.2-1 08.2 Average Recovery (%) 1 03.7 1 :4 Range (%) 1 00. 7-108.4 Average Recovery (%) 1 1 1 .1 1 :8 Range (%) 1 00.7-1 1 7.2 Average Recovery (%) 1 1 4.7 1 :1 6 Range (%) 1 08.8-1 1 7.7 Serum 91 .9 84.5-99.8 97.3 88.8-107.2 92.8 84.9-1 03.0 93.3 81 .4-1 05.8 Citrate plasma 96.4 88.8-103.9 95.9 88.3-1 01 .0 92.6 81 .2-99.9 99.2 9 1 . 1 -1 07.0 To assess the linearity of the assay, samples spiked with high concentrations of human IL-15 were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay. T e l : + l 8 0 0 -8 1 0 - 0 8 1 6 • W e b : w w w . a c ro b i o sy s t e m s . c o m E m a i l : o rd e r@ a c r o b i o s y s t e m s . c o m ► Intra-Assay Statistics Sample 2 3 Number of Replicate 20 20 20 Mean (pg/ml) 95.805 27.934 8.71 8 SD 5.227 1 .629 0. 669 CV (%) 5.5 5.8 7.7 Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision, Intra-Assay Precision CV < 1 0%. ► Inter-Assay Statistics Sample 2 3 Number of Replicate 3 3 3 Mean (pg/ml) 93.999 28.455 9. 573 SD 5.629 1 .900 0.783 CV (%) 6.0 6.7 8.2 Three samples of known concentration were tested in three separate assays to assess inter-assay precision, Inter-Assay Precision CV< 1 0% . ► Recovery Sample Type Serum(n= 5) Average % Recovery 86.3% Range 80.9-97.7% Three parts of blank serum were added with different concentrations of human IL-1 5 , and the serum without human I L- 1 5 was used as background to calculate the recovery rate. The range of the recovery rate is 80. 9-97. 7%, and the average recovery is 86.3%. [ resDetect™ Antibiotic Residue Detection Kit ACROBiosystems, through rigorous methodological validation, has independently developed the resDetect™ Kanamycin ELISA Kit and resDetect™ Gentamicin ELISA Kit. The calibration standards for these kits trace back to regulatory standards (Kanamycin 130556; Gentamicin 130326). They are suitable for the quantitative determination of residual antibiotics in plasmid DNA raw materials and proteins for cell and gene therapy (CGT), vaccines, and other biopharmaceuticals. ■ Features i;6' High specificity: No cross-reactivity with ampicillin, tetracycline, and chloramphenicol. i;6' Convenient and efficient: Simple operation, using a one-step method, results can be obtained in just 1 hour and 20 minutes. i;6' Excellent performance: Good linearity of the standard curve, high accuracy, and good repeatability. i;6' Good stability: Small batch-to-batch variation, long shelf life. ■ Product List Cat. No . RES-A004 RES-A025 Description resDetect'" Kanamycin ELI SA Kit resDetect'" Gentamicin ELI SA Kit T e l : +l 8 0 0 - 8 1 0 - 0 8 1 6 • W e b : w w w . a c r o b i o s y s t e m s . c o m E m a i l : o rd e r @ a c r o b i o s y s t e m s . c o m Size 9 6 T 9 6 T ■ Features Cat. No. Detection range Recovery Precision Specificity ► Standard Curve * RES-A004 Stand ard Cone. (ng/ml) 40.5 1 3. 5 4.5 1 . 5 0.5 O D 450-1 0.203 0 . 3 52 0 .643 1 .2 5 8 2.1 9 5 0.5~40.5ng/ m l 70%~130% < 15% RES-A004 No cross-reactivity with ampicillin, tetracycline, ch/oramphenico/; No cross-reactivity with E. coli HCP, DNA. O D 450-2 Average 0 . 1 8 5 0 . 1 94 + I- 0 . 345 0. 349 + 0 . 6 3 5 0 . 6 3 9 I- 1 .207 1 .2 3 3 2 . 1 6 5 2 . 1 8 2.5 2.0 1 .5 1 .0 0.5 RES-A025 0.25~8 ng/ m l 70%~130% < 15% No cross-reactivity with ampicillin, tetracycline, chloramphenicol; No cross-reactivity with E. coli HCP, DNA. 10 20 30 40 50 Kanamydn Cone. (ng/ml) For each experiment, a standard curve needs to be set for each micro-plate, and the specific OD value may vary depending on different laboratories, testers, or equipments. The following example data is for reference only. * RES-A025 Stand a rd C o n e . (ng/ m l) OD 450-1 O D 450-2 Average 0.5 8 0. 1 33 0.1 53 0. 1 43 + I- 4 0.202 0.222 0.21 2 21 0.0 + t 2 0.354 0.388 0.371 "' I- 3 -0.5 0.625 0.704 0.665 + I- 0.5 1 .287 1 .443 1 .365 + I- -1.0 0.25 2.223 2.271 2.247 0 4 8 1 0 Gentamlcin Cone. (ng/mL) Serial dilutions of gentamicin (from 8 nglmL to 0.25 nglmL) was added into gentamicin: anti-gentamicin binding reactions. The assay was performed according to the protocol dscribed below. Background was subtracted from data points prior to log transformation and curve fitting. ► resDetect ™ Kanamycin ELISA Kit * Validated results showed no significant cross-reactivity when separately adding S00μg/ml ampicillin, tetracycline, and chloramphenicol in the sample diluent. Additionally, when adding 2000ng/ml E. coli host protein, 200ng/ml E. coli host DNA, and 50ng/μL plasmid DNA in the diluent, the recovery rates for all three samples were between 70-1 30%. Cross Reactant Kanamycin (500ug/ml) Ampicillin (500ug/ml) Tetracycline (500ug/ml) Chloramphenicol (500ug/ml) Cross-reactivity Cross-reactivity 1 00% < 1 % < 1 % < 1 % Tel: + l 800-810-0816 • Web: www.acrobiosystems.com Email: order@acrobiosystems.com I nterference Interference Factor E.coli HCP Cone. (2000ng/ml) E.coli HCD Cone. (200ng/ml) Plasmid DNA Cone. (50ng/μL) Sample Conc.(ng/ml) 30 Detected Sample Cone. (ng/ml) 27.79 Recovery Rate (%) 92 4.5 5.47 1 1 6 ► resDetect ™ Gentamicin ELISA Kit 1 . 1 7 1 1 7 30 32.42 1 08 4.5 4.83 1 07 1 .22 1 22 30 39.59 1 30 4.5 6.23 1 1 8 1 .34 1 27 * Validated results showed no significant cross-reactivity when separately adding S00μg/ml ampicillin, tetracycline, and chloramphenicol in the sample diluent. Additionally, when adding 2000ng/ml E. coli host protein, 200ng/ml E. coli host DNA, and 1 00ng/μL plasmid DNA in the diluent, the recovery rates for all three samples were between 70-1 30%. Cross Reactant Gentamicin (500ug/ml) Ampici llin (500ug/ml) Tetracycline (500ug/ml) Chloramphenicol (500ug/ml) Cross-reactivity I nterference Cross-reactivity 1 00% < 1 % < 1 % < 1 % Cross Reactant E.coli HCP Cone. (2000ng/ml) E.coli HCD Cone. (200ng/ml) Plasmid DNA Cone. (1 00ng/μL) Sample Conc.(ng/ml) Detected Sample Cone. (ng/ml) Recovery Rate (%) 5 4.57 91 2 2 1 00 0.5 0.4 80 5 5.09 1 02 2 2.2 1 1 0 0.5 0.58 1 1 6 [ resDetect™ GENIUS™ Nuclease ELISA Kit 5 6.06 1 21 2 2.4 1 20 0.5 0.62 1 25 In the process of treating bioproducts with nucleases, trace amounts of nucleases may remain. Since the broad-spectrum nuclease is classified as a contaminant, these trace residues impact on the subsequent application of bioproducts and may cause toxicity or immune reactions. Therefore, precise detection of nuclease residue is a crucial factor in ensuring the activity and safety of related bioproducts. Our GENIUS™Nuclease ELISA Kit (Catalog number: CRS-A016), a highly sensitive and specific assay for the accurate detection of broad-spectrum nuclease residue, addressing your concerns regarding nuclease residue. ■ Featu res 􀀶 High sensitivity up to 2.733pg/ml, allowing precise quantification of trace residues of nucleases in culture medium supernatants. 􀀶 Strong adaptability, compatible not only with ACRO's own broad-spectrum nuclease but also with many enzyme products on the market. 􀀶 Stable product performance, with intra- and inter-batch precision below 10%. 􀀶 Complete methodological validation reports are available for free. 􀀶 Self-produced raw materials, high production yield, and short delivery time. Tel: +l 800-810-0816 • Web: www.acrobiosystems.com Email: order@acrobiosystems.com ■ Data Q5 R2=0.􀁐999 • ···· ···· ··· • •1 .,<-- - --- - · - · _ ____ ___ · · - --- --- -- - -- · --- - · I For each experiment, a standard curve needs to be set for each micro-plate, and the specific OD value may vary depending on different laboratories, testers, or equipments. The following example data is for reference only. 500 7000 1500 Conc.(pg/ml) 21») 2500 3.5 3 0 2.5 2.0 L5 1.0 0.5 0 0 -+-ACRO ---ven dor- 1 􀀮vendor-2 3000 6000 9000 12000 Sample Conc,lpg/ml) GENIUS™ Nuclease ELISA Kit (Residue Testing) (Cat No. CRS-A016) can detect the nuclease from different manufacturers with similar sensitivity. 4 2 0 . 5%BSA E coli c ell supernatant CHO c ell supernatant HEK.293 c ell supernatant 0 0.01 0,. 1 1 0 1 0 0 Nuclease Cone. (ng,lml) GENIUS™ Nuclease ELISA Kit (Residue Testing) (Cat No. CRS-A0 1 6) can detect the nuclease in different culture supernatant with similar curve and sensitivity. 0.120 0.100 0.080 0.060 0 0.040 0.020 0.000 Positive Negative E.coli CHO H EK2 93 1 %BSA GENIUS ™ Nuclease ELISA Kit (Residue Testing) (Cat No. CRS-A016) can detect the nuclease from different culture supernatant and was free from the matrix effects. *means a significant difference compared with the negative, NS means no significant difference with the negative. Tel: +l 800-810-0816 • Web: www.acrobiosystems.com Email: order@acrobiosystems.com [ resDetect TM DNase Activity Assay Kit & RNase Activity Assay Kill Through rigorous methodological validation, we have independently developed DNase and RNase residue detection kits based on the fluorescence probe method. These kits can be used to determine whether the environment, consumables, raw materials, etc., are contaminated with DNase and RNase. They are designed to detect the residual levels of DNase and RNase in bioproducts, enabling the development of appropriate clearance solutions to meet research and production needs. ■ Detection Principle Fluorescence probe-based DNase and RNase residual activity detection relies on fluorescence-labeled RNA and DNA substrates. When the sample does not contain RNase and DNase activity, the substrate remains stable, and the fluorescent and quenching moieties are close together. Due to the principle of fluorescence resonance energy transfer, no fluorescence signal is generated. When the sample contains RNase and DNase activity, the substrate degrades, causing the fluorescent and quenching moieties to move away from each other, resulting in an gradually enhanced fluorescence signal. The rate of increase in fluorescence signal is positively correlated with the quantity and activity of the enzymes. Using a fluorescence microplate reader at wavelengths ex/em=490/520nm (RNase) and 5351565nm (DNase), one can determine whether the sample is contaminated with DNase/RNase. ■ Features Fluorescent Substrate Fluorescent Substrate Fluo rescent Substrate Fluorescent Substrate No fluorescent signal Have fluorescent signal No DNase activity No fluorescent signal Have DNase activity 􀀶 Designed based on the principle of enzyme activity, suitable for detecting residual RNase or DNase of various types. 􀀶 High sensitivity, with a detection limit as low as 3.9*10-SU (DNase)/0.03pg (RNase). 􀀶 Strict quality control, ensuring sensitivity, inter-batch/intra-batch variation, accuracy, freeze-thaw stability, etc. 􀀶 Comprehensive method validation in accordance with ICH Q2(R2) guidelines. 􀀶 Simple and fast to use, with the ability to complete testing for over 80 samples within 30 minutes. 􀀶 Verified recovery rate, eliminating interference from glycerol stabilizers in the original enzyme. 􀀶 Can be used in combination, allowing simultaneous detection of RNase or DNase residue without interference. ■ Product List Cat. No. ASE-A001 ASE-A002 ■ Product Data Product Description RNase Activity Assay Kit (Fluorescence) DNase Activity Assay Kit (Fluorescence) Cut-off Criteria: Using the enzyme-linked immunosorbent assay (ELISA) method, if the fluorescence ratio between the test sample and the negative control sample is <2, it is determined as no residual DNase/RNase. Otherwise, if the ratio is <::2, it is determined as having residual DNase/RNase. ► RNase Activity Assay Kit (Fluorescence) (ASE-A00 1 ) * The validated sensitivity of this reagent can reach 0.031 25pg. 60000 time (min) -+- 2pg - 1 pg - 0.5pg - 0.25pg - 0 . 1 25pg -+- 0.0625pg - 0.03125pg - 0pg Add 90 μL of the working RNase Substrate solution to each 96-well plate, and add 10 μL of RNase A standards (0-200pg/ml *10μL / well = 0-2pg/well), incubate the plate in the fluorometer (BMG CLARIOstar) collecting real-time data at 1 minute intervals for 30 minutes at 37°C using the settings described in this section. The RNase Activity Assay can be evaluated in rigorous kinetic terms using real-time data. Tel: +l 800-810-0816 • Web: www.acrobiosystems.com Email: order@acrobiosystems.com * The intra-assay variability of RNase Activity Assay Kit (Fluorescence) (Cat. NO. ASE-A001 ) is less than 1 0%, inter-assay variability is less than 1 5%, and the precision meets standards set in regulatory guidelines. Intra-Assay Statistics Sample 2 3 4 5 6 7 N u m ber of Replicate 8 8 8 8 8 8 8 Mea n RFU 3506 5964 9831 1 6862 27758 41779 56643 Standard Deviation 1 30 1 92 291 532 1 80 783 412 Coefficient of Variation (96) 3.7 3.4 3.0 3.2 0.6 1 .9 0.7 Inter-Assay Statistics Sa m p le 2 3 4 5 6 7 Number of Re plicate 8 8 8 8 8 8 8 Mea n RFU 3858 6132 1 0503 1 7609 28071 41 161 53533 Standard Deviation 579 769 1 495 2307 3341 4203 4249 Coefficient of Variation (96) 15 12.5 1 4.2 1 3 .1 1 1 .9 1 0.2 7.9 * Verified, the recovery rate of RNase Activity Assay Kit (Fluorescence) (Cat. No. ASE-A001 ) is within the range of 80% to 1 20%. 5.5 μg/ml of Pyrophosphatase 1 % of (50mM Tris-HCI and 50% 2 μg/ml Thermostable System I norganic Pyrophosphatase 1 xReaction Buffer (n=2) (n=2) Glycerol) (n=2) (n=2) Sample weight. (pg) Calculated Ave Calculated Ave Calculated Ave Calculated Ave weight. (pg) % RE weight. (pg) % RE weight. (pg) % RE weight. (pg) % RE Sample 1 1.5 1 .4252 95 1.3319 89 1 .3375 89 1 .4193 95 Sample 2 0.2 0.1691 85 0.1669 83 0.1 683 84 0.1839 92 Sample 3 0.05 0.0437 87 0.0461 92 0.0442 88 0.0472 94 * After three freeze-thaw cycles of the probe, the performance meets the requirements, and there is no decrease in sensitivity. The CV is less than 1 0%. 80000 60000 􀀯 40000 20000 ..... No Freezing and thawing ..... Freezing and thawing 1 times ..... Freezing and thawing 2 times ..... Freezing and thawing 3 times 0-1-􀀰􀀱􀀲....--􀀳􀀴􀀵...,.-􀀶􀀷􀀸 ........ 0.01 0.1 1 RNase A (pg) T e l : + l 8 0 0 - 8 1 0 - 0 8 1 6 • W e b : www . a c r o b i o s y s t e m s . c o m 10 E m a i l : o r d e r @ a c r o b i o s ys t e m s . c o m * No cross-interference: Verified using DNase Activity Assay Kit (Fluorescence) (Cat. No. ASE-A002) and RNase Activity Assay Kit (Fluorescence) (Cat. No. ASE-A001 ) to detect the residual nucleases in the same sample, and no cross-interference was observed. 50000 -40000 ::::1 30000 􀀳 20000 10000 10 20 time (min) JO 40000 -+- DNase SubslnHe ]0000 􀀴 20000 10000 ► Competitor Evaluation Data DNst,e l OU RN.1􀀕 A:2pg time- (min) - RNas.e Subs1raIe 40000 30000 􀀠 20000 1 0000 - ONas.e Substrale -+- RNase Substrale time (rnin) * The sensitivity of this kit can reach 0.031 25pg, comparable to the performance of the imported brand Competitor T, and superior to most other brands on the market. 60000 40000 => =>LL. c,: 20000 6000 ACRO 1 0 20 time (min) Competitor H 10 20 time (min) - 2pg - 1pg -- 0.5pg ..... 0.25pg - 0.125pg - 0.0625pg ..... 0.03125pg - 0pg 30 -+- 0.2 pg -+- 0.1 pg - 0.05 pg -+- 0.025 pg 0 pg 30 Tel: +l 800-810-0816 • Web: www.acrobiosystems.com Competitor T 60000 - 2pg ......... 1pg 40000 -- 0.5pg => -+- 0. 25pg LL. -+- 0.125pg c,: 20000 ..... 0. 0625pg -+- 0.03125pg -+- 0pg 10 20 30 time (min) Competitor B 40000 20pg 30000 1 0pg 5pg => -+- 2 .5pg 􀀏 20000 1 . 25pg 0.625pg 10000 -+- 0pg 10 20 30 time (min) Email: order@acrobiosystems.com A BIOSYSTEMS • I w􀀂 Pv-o􀀅 􀀆 I􀀇 c r o A 􀂄VvC.,e, B l/0--Vv\.-e,,oLitC/i.,,􀀕 C o pyri g ht State m e nt ' ' This material is copyrighted by the Company. All rig hts in this material are reserved by the Company. Unless otherwise indicated in writing, all m aterial i n this materi a l is copyrig hted by the C o m p a n y . N o p a rt of t h i s m ate r i a l m a y b e copied , photocopied or reproduced i n any form or red istri buted to any other person or u sed in any other manner which i nfringes the Company's copyrig ht without the prior written authorisation of the Company. 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