Detection of recombinant lysostaphin using antibody sandwish enzyme-linked immunoadsorbent assay.
- Author:
Qing-Shan HUANG
1
;
Ji-En ZHANG
;
Hong-Yu WU
;
Yun-Jie MO
Author Information
1. State Key Laboratory of Genetic Engineering, Insititute of Genetics, School of Life Science, Fudan University, Shanghai 200433, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Monoclonal;
immunology;
Blotting, Western;
Enzyme Stability;
Enzyme-Linked Immunosorbent Assay;
methods;
Humans;
Immune Sera;
immunology;
Lysostaphin;
blood;
immunology;
metabolism;
Male;
Rabbits;
Recombinant Proteins;
immunology;
metabolism;
Reproducibility of Results;
Temperature
- From:
Chinese Journal of Biotechnology
2007;23(1):117-121
- CountryChina
- Language:Chinese
-
Abstract:
The double-antibody-sandwich enzyme-linked immunoadsorbent assay (ELISA) for detection of rLysostaphin in humans had been developed and established through this study. rLysostaphin of high purity ( > 95 % ) produced in Shanghai Hi-Tech United Bio-Technological Research & Development Co., Ltd (SHUBRD) was used to produce a rabbit anti-rLysostaphin polyclonal antibody. The standard curve of rLysostaphin polyclonal antibody that was constructed showed that the lowest range of detection was found at 0. 98 ng of rLysostaphin/mL, and the curve exhibited linearity preferably from 0. 98 to 500 ng of rLysostaphin/mL. When three serum samples of the same batch were assayed for 6 replicates, and more 3 samples from different batches for 6 replicates, the average intra-assay and inter-assay coefficient variances ( CV) were 6. 4% and 6. 5%, respectively. The relative recovery rate was 98.6% when quantitative standard antigens were added to the serum. The present method for detection of rLysostaphin in serum is specific, highly sensitive, highly precise, and exhibited a low CV and will be helpful in the further study of rLysostaphin pharmacokinetics and holds promise in clinical applications.
