[Detection of recombinant lysostaphin using antibody sandwish enzyme-linked immunoadsorbent assay].
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.
Huang
Qing-Shan
QS
State Key Laboratory of Genetic Engineering, Insititute of Genetics, School of Life Science, Fudan University, Shanghai 200433, China.
Zhang
Ji-En
JE
Wu
Hong-Yu
HY
Mo
Yun-Jie
YJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western; D004795:Enzyme Stability; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007106:Immune Sera / Q000276:immunology*; D008248:Lysostaphin / Q000097:blood / Q000276:immunology* / Q000378:metabolism; D008297:Male; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D015203:Reproducibility of Results; D013696:Temperature
D000911:Antibodies, Monoclonal; D007106:Immune Sera; D011994:Recombinant Proteins; D008248:Lysostaphin