| Protein A ELISA Kit Protein A affinity chromatography is widely used for antibody purification. However, the leakage of Protein A, though covalently conjugated to chromatography supports, posts a big problem for antibody based pharmaceutical industry. And it also probably causes false results in immunological assays if antibody product is contaminated with Protein A.
GenScript Protein A ELISA Kit is developed for accurate quantification of the leakage of native Protein A, recombinant Protein A variants and MabSelect SuRe™ in the presence or absence of IgG antibodies. The kit offers users in pharmaceutical industry a sensitive assay to measure mini amount of Protein A in therapeutic antibody products. It also facilitates Protein A resin manufacturers in monitoring leaching characteristics of the resins under specific conditions. | Key Features: - Accurate detection of 20 pg/ml Protein A and 50 pg/ml MabSelect SuRe™ in antibody preps.
- Broad-range recognition of native Protein A, recombinant Protein A variants and MabSelect SuRe™
- Low level deviation - Intra-assay <5%, Inter-assay <10%
- High accuracy/recovery within 85 -115%
- Ready-to-use ELISA plate with fast & simple samples preparation
- Time-saving protocol - complete assay in less than 2 hours
- Exceptional lot-to-lot consistency ensured by high-quality monoclonal antibodies
- Wide applications for the quantification of Protein A contaminant and leaching
- Specifications
- Examples
- Related Products
Components | Quantity | 96-well plate coated with anti-Protein A mAb | 1 plate (8 wells x 12 strips) | Biotin conjugated anti-Protein A mAb | 12 ml | Streptavidin-HRP | 12 ml | Protein A Standard (10 μg/ml) | 1 ml | MabSelect SuRe™ Protein A Standard (10 μg/ml) | 1 ml | 20 × Wash Solution | 40 ml | One-Solution Microwell TMB Substrate | 12 ml | Stop Solution | 6 ml | | All components should be stored at 2-8 °C. | |  | Typical Assay Data |  | Conc. of Protein A (ng/ml) | OD 450 | Duplicate 1 | Duplicate 2 | Average | Adjusted Average | 3.20 | 1.711 | 1.829 | 1.770 | 1.717 | 1.60 | 0.849 | 0.871 | 0.860 | 0.807 | 0.80 | 0.461 | 0.468 | 0.465 | 0.412 | 0.40 | 0.247 | 0.257 | 0.252 | 0.199 | 0.20 | 0.163 | 0.164 | 0.164 | 0.111 | 0.10 | 0.141 | 0.149 | 0.145 | 0.092 | 0.00 | 0.077 | 0.076 | 0.077 | 0.024 | Blank | 0.052 | 0.054 | 0.053 | | |
Figure 1. Standard curve generated using GenScript Protein A ELISA Kit (L00430). | | | Precision | intra-assay | inter-assay | # of tests | Meanng/ml | CV% | # of assays | Meanng/ml | CV% | 12 | 1.6 | 1.60 | 8 | 1.6 | 3.70 | 12 | 0.4 | 4.05 | 8 | 0.4 | 5.42 | 12 | 0.1 | 4.56 | 8 | 0.1 | 7.59 | | Precision is defined as the percent coefficient of variation (%CV). The data above shows CV value of both intra and inter-assay for 3 control samples in low, mid and high concentration range. Intra-assay Precision CV<5% and Inter-assay Precision CV<10% | | | Recovery |
Sample Matrix | Protein A Added (ng/ml) | % Recovery | 6.20 mg/ml human IgG | 1.50 | 85.01 | 1.19 mg/ml customer Ab | 1.00 | 98.07 | 1.13 mg/ml human IgG | 1.20 | 108.96 | 1.00 mg/ml human IgG | 2.50 | 100.01 | | The assay was performed by spiking the Protein A standards provided in this kit into the IgG samples. The recovery rates are within 85% to 115%. | | | | |
