Production and characterization of anti-estrone monoclonal antibody / 生物医学与环境科学(英文)
Biomedical and Environmental Sciences
;
(12): 103-112, 2002.
Article
in English
| WPRIM
| ID: wpr-264327
ABSTRACT
<p><b>OBJECTIVE</b>Determination of estrone (E1) levels has a significant meaning in evaluating physiological effect and diagnosing some diseases. In order to detect free E1 in biological fluids, a monoclonal antibody specific for E1 was prepared after the complete antigen of E1 was synthesized. The purified monoclonal antibody was fully characterized for later immunoassay.</p><p><b>METHODS</b>3-O-carboxymethyl ether derivative of E1 was synthesized and in turn coupled to bovine serum albumin (BSA) to form complete antigen E1-BSA. A monoclonal antibody (McAb) specific for E1 was produced both in vitro and in vivo by a hybridoma anti-E1. Anti-E1 was prepared by fusion of SP2/0 murine myeloma cells with spleen cells isolated from immunized BALB/c mouse. The McAb was characterized by enzyme-linked immunosorbent assay (ELISA), SDS-PAGE and Western-blotting. The specificity of the immunoassay was investigated by determining the cross-reactions of E1 analogs when free E1 was detected by competitive indirect enzyme-linked immunosorbent assay (CI-ELISA).</p><p><b>RESULTS</b>Analysis revealed that anti-E1 McAb (E1-McAb) was of the IgG1 type, the molecular weight of E1-McAb was 164,000 daltons. The affinity constant of E1-McAb with coated complete antigen was 8.2 x 10(8) L/mol. The linear range for free E1 determined by CI-ELISA was 10 pg/mL-10 ng/mL. The detection limit was 21.4 pg/mL (defined as twice the standard deviation of the blank).</p><p><b>CONCLUSION</b>The CI-ELISA developed with E1-McAb was both sensitive and specific. The prepared E1-McAb can be used in some immunoassays.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Enzyme-Linked Immunosorbent Assay
/
Blotting, Western
/
Cross Reactions
/
Electrophoresis, Polyacrylamide Gel
/
Allergy and Immunology
/
Estrone
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Hybridomas
/
Mice, Inbred BALB C
/
Antibodies, Monoclonal
/
Antibody Affinity
Limits:
Animals
Language:
English
Journal:
Biomedical and Environmental Sciences
Year:
2002
Type:
Article
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