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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.
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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 |
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All components should be stored at 2-8 °C. |
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Typical Assay Data |
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Conc. of
Protein A (ng/ml)
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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 |
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Figure 1. Standard curve generated using GenScript Protein A ELISA Kit (L00430). |
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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 |
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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% |
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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 |
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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%. |
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