Volume 15 Issue 2
Jun.  2002
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WANG YONG-CHENG, GUO ZHEN-QUAN, LI Yuan-zong, CHANG Wen-bao. Production and Characterization of Anti-estrone Monoclonal Antibody[J]. Biomedical and Environmental Sciences, 2002, 15(2): 103-112.
Citation: WANG YONG-CHENG, GUO ZHEN-QUAN, LI Yuan-zong, CHANG Wen-bao. Production and Characterization of Anti-estrone Monoclonal Antibody[J]. Biomedical and Environmental Sciences, 2002, 15(2): 103-112.

Production and Characterization of Anti-estrone Monoclonal Antibody

Funds:  国家自然科学基金(Grant 20075001)
  • Objective 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. Methods 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). Results 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′108L/mol. The linear range for free E1 determined by CI-ELISA was 10pg/mL~10ng/mL. The detection limit was 21.4 pg/mL (defined as twice the standard deviation of the blank). Conclusion The CI-ELISA developed with E1-McAb was both sensitive and specific. The prepared E1-McAb can be used in some immunoassays.
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Production and Characterization of Anti-estrone Monoclonal Antibody

Funds:  国家自然科学基金(Grant 20075001)

Abstract: Objective 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. Methods 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). Results 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′108L/mol. The linear range for free E1 determined by CI-ELISA was 10pg/mL~10ng/mL. The detection limit was 21.4 pg/mL (defined as twice the standard deviation of the blank). Conclusion The CI-ELISA developed with E1-McAb was both sensitive and specific. The prepared E1-McAb can be used in some immunoassays.

WANG YONG-CHENG, GUO ZHEN-QUAN, LI Yuan-zong, CHANG Wen-bao. Production and Characterization of Anti-estrone Monoclonal Antibody[J]. Biomedical and Environmental Sciences, 2002, 15(2): 103-112.
Citation: WANG YONG-CHENG, GUO ZHEN-QUAN, LI Yuan-zong, CHANG Wen-bao. Production and Characterization of Anti-estrone Monoclonal Antibody[J]. Biomedical and Environmental Sciences, 2002, 15(2): 103-112.

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