Development of ELISA to estimate thymosin alpha1, the N terminus of prothymosin alpha, in human tumors.
We reported that tumor content of prothymosin alpha (ProT alpha) is a proliferation index of human breast tumors that might be used to identify patients at high risk for distant metastasis (Dominguez et al., Eur J Cancer 1993; 29A:893-7). In that study ProT alpha concentrations were measured by a RIA; here we present an alternative nonisotopic assay that could be used in a standard clinical laboratory. Main features of the ELISA are: (a) A recombinant fusion protein glutathione S-transferase (GST)-human ProT alpha was used to coat the microtiter plates; (b) we used a polyclonal antiserum raised in rabbits that detects thymosin alpha1, the NH2-terminal fragment of ProT alpha; (c) it is as sensitive as the RIA; (d) it is faster than the RIA. ProT alpha concentrations in various human tumors (skin, esophagus, colorectal, and breast) as assessed by ELISA were comparable with, although twofold greater than, the values previously estimated by RIA.
Loidi
L
L
Departamento de Fisiología, Facultad de Medicina, Universidad de Santiago, Spain.
Vidal
A
A
Zalvide
J B
JB
Puente
J L
JL
Reyes
F
F
Domínguez
F
F
D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000706:statistics & numerical data; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D009369:Neoplasms / Q000737:chemistry*; D010446:Peptide Fragments / Q000032:analysis; D011498:Protein Precursors / Q000032:analysis*; D012044:Regression Analysis; D012680:Sensitivity and Specificity; D000077596:Thymalfasin; D013947:Thymosin / Q000031:analogs & derivatives* / Q000032:analysis
D007106:Immune Sera; D010446:Peptide Fragments; D011498:Protein Precursors; C040782:prothymosin alpha; D013947:Thymosin; D000077596:Thymalfasin