Development of a sandwich enzyme-linked immunosorbent assay (ELISA) for determining of bovine serum albumin (BSA) in trivalent measles-mump-rubella (MMR) vaccines

Abstract

A sandwich enzyme-linked immunosorbent assay (ELISA), using polyclonal antibody, was developed and compared with the commercial kit for detecting and estimating of BSA content in Measles-Mump-Rubella (MMR) vaccine samples in detection limit of nanogram level. The test depends on the capturing and detecting of BSA antigen by the polyclonal antibody. Initially, a detection range of 0-64 ng/ml was established, could be used for estimation of BSA content according to WHO requirement (50 ng/ml) in MMR vaccines. Comparative analysis of the test results for 85 MMR vaccine samples obtained with the commercial kit gave a sensitivity of 58.8% and a specificity of 97%. A high correlation (r = 0.94) was observed between BSA sandwich ELISA and commercial kit for BSA content in MMR samples. However, variations in values also were observed for the two assays. These variations may have been due to difference of upper limit of detection range of BSA content in commercial kit (32 ng/ml) and new sandwich ELISA (64 ng/ml) as well as the use of a different polyclonal antibody. In concerning the cutoff value for the WHO requirement and employment of standard solution of 64 ng/ml in developing assay, it would be adequate to use this test for assessing BSA content in viral vaccines same as MMR vaccines.

Highly Sensitive Chemiluminescent Analysis of Residual Bovine Serum Albumin (BSA) Based on a Pair of Specific Monoclonal Antibodies and Peroxyoxalate–glyoxaline–PHPPA Dimer Chemiluminescent System in Vaccines

“…Among all kinds of vaccines, many are derived from cell-culture-grown viruses [17]. Calf sera are often taken in the culture medium for culturing the cells which often are used to produce vaccines [18,19]. BSA is a major component of calf sera, which is frequently used for cell culture.…”

The establishment of a highly sensitive ELISA for detecting bovine serum albumin (BSA) based on a specific pair of monoclonal antibodies (mAb) and its application in vaccine quality control

“…Some viral vaccines are purified from cell cultures that are infected with viruses, such as the measles vaccine, and hepatitis B vaccine contained a certain concentration of calf sera. 3 Many observations in humans have called attention to the possible sensitizing property of this kind of vaccine. With the increasing number of reports about the diseases caused by BSA, [4][5][6][7] the problem of residual bovine serum proteins, especially BSA, in viral vaccines has drawn more and more attention.…”

“…With the linear part of the curve for the standard optimized for low concentrations and with the limit between negative and positive results set at the mean plus 2 SD for the concentrations of and expensive, 12,13 and the sandwich ELISA. 3,10 Among these methods, the sandwich ELISA was found to be advantageous for specifically detecting native BSA in biological samples in the presence of other proteins. The key issue in this assay is the correct antibodies used to establish the ELISA.…”

Development of an ELISA for Evaluation of Swab Recovery Efficiencies of Bovine Serum Albumin

“…This study presents a BSA sandwich ELISA with a quantification range from 1.32–40 ng/mL for quantification of BSA from swab samples ( figure 2 ). The ELISA has comparably sensitivity to other BSA sandwich ELISAs, both commercial assays and assays described in the literature [10] , [11] , [12] . The benefits of using the described ELISA are that the setup is simple and all components are commercially available.…”

“…These factors affect chemical and physical properties such as solubility and sensitivity to denaturation of the protein [8] . Several methods have been developed to detect BSA, including counter current electrophoresis, spectrofluorimetric, label-less immunosensors, immunodiffusion, regular and sandwich ELISA [9] , [10] , [11] , [12] , [13] . For this study we needed a simple and easy detection method in order to quantify BSA in the context of dispersal and swab experiments.…”