Protein A ELISA Kit

Overview

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. Moreover, 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 the pharmaceutical industry a sensitive assay to measure mini amounts 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%

Time-saving protocol - complete assay in less than 2 hours

Ready-to-use ELISA plate with fast & simple samples preparation

Exceptional lot-to-lot consistency ensured by high-quality monoclonal antibodies

Wide applications for the quantification of Protein A contaminant and leaching

Specifications

Examples

• 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%.

Related Products