Development and validation of an ELISA method for detection of growth arrest specific 6 (GAS6) protein in human plasma.
Gas6 protein is possibly involved in human diseases, but a validated plasma assay is lacking. So, we developed a sandwich enzyme-linked immunosorbent assay (ELISA) method using commercially available reagents. An appropriate plasma-based matrix was prepared to optimize the assay. The ELISA method showed inter- and intra-assay coefficients of variation lower than 15%. Recoveries all fell within 15% of expected values. Plasma Gas6 concentration in 61 healthy donors was 20.3+/-3.8 ng/mL. Our assay meets FDA requirements for precision and accuracy for the validation of bioanalytical methods and it is suitable for research or diagnostic purposes.
Alciato
Federica
F
Department of Clinical and Experimental Medicine, Università del Piemonte Orientale, Novara, Italy.
Sainaghi
Pier Paolo
PP
Castello
Luigi
L
Bergamasco
Luca
L
Carnieletto
Stefania
S
Avanzi
Gian Carlo
GC
D000328:Adult; D000368:Aged; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards; D005260:Female; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000097:blood*; D008297:Male; D008875:Middle Aged; D010949:Plasma / Q000378:metabolism; D012680:Sensitivity and Specificity; D014481:United States; D014486:United States Food and Drug Administration; D000099255:Growth Arrest-Specific Protein 6
D036341:Intercellular Signaling Peptides and Proteins; D000099255:Growth Arrest-Specific Protein 6