Monoclonal antibody based ELISAs for measurement of activins in biological fluids.
Two sensitive monoclonal antibody (MAb)-based enzyme-linked immunosorbent assays (ELISAs), one for activin A (homodimer of beta A subunits) and one for activin B (homodimer of beta B subunits) in plasma have been developed. The activin A ELISA had an effective range of 0.2-50 ng/ml while the activin B ELISA's range was 0.1-25 ng/ml in human serum. Both ELISAs were specific with < 0.01% cross-reactivity with related hormones and follistatin (an activin binding protein), however the presence of recombinant human follistatin caused a decrease in measured level of activin A and B spiked human samples. The assay was linear across the standard curve range with intra- and interassay coefficients of variation were less than 15%. The level of activins in female serum range from 0.3 to 10.4 ng/ml. In summary, we have developed a reliable, convenient and rapid MAb-based enzyme immunoassay for determination of activin A and B levels in human serum which are also applicable for buffer, mouse and monkey serum matrices. This assay will be useful for studying the regulation and role of activin A and B in health and disease.
Wong
W L
WL
Department of Medicinal and Analytical Chemistry, Genentech Inc., So. San Francisco, CA 94080.
Garg
S J
SJ
Woodruff
T
T
Bald
L
L
Fendly
B
B
Lofgren
J A
JA
D028341:Activins; D000276:Adjuvants, Immunologic / Q000032:analysis*; D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006133:Growth Substances / Q000032:analysis*; D006801:Humans; D007265:Inhibins / Q000032:analysis*; D051379:Mice; D008807:Mice, Inbred BALB C; D009842:Oligopeptides; D010455:Peptides / Q000032:analysis*; D011994:Recombinant Proteins / Q000032:analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000276:Adjuvants, Immunologic; D000911:Antibodies, Monoclonal; D006133:Growth Substances; D009842:Oligopeptides; D010455:Peptides; D011994:Recombinant Proteins; C044147:activin B; D028341:Activins; C048617:myelopeptides; D007265:Inhibins