Development of an ELISA method for detecting immune complexes between tissue-nonspecific alkaline phosphatase and immunoglobulin G.

Development of an ELISA method for detecting immune complexes between tissue-nonspecific alkaline phosphatase and immunoglobulin G. Research Abstract Details 

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Development of an ELISA method for detecting immune complexes between tissue-nonspecific alkaline phosphatase and immunoglobulin G. Abstract Text:

Kazuo Hocchi,Tatsuya Ohashi,Toshihide Miura,Kumiko Sasagawa,Yasuhito Sato,Fumio Nomura,Takeshi Tomonaga,Masahiko Sunaga,Ryo Kojima,Katsuhiro Katayama,Toshiyuki Kato,Toyoji Sato,Tsugikazu Komoda,Kimimitsu Oda,Kazuo Hocchi,Tatsuya Ohashi,Toshihide Miura,Kumiko Sasagawa,Yasuhito Sato,Fumio Nomura,Takeshi Tomonaga,Masahiko Sunaga,Ryo Kojima,Katsuhiro Katayama,Toshiyuki Kato,Toyoji Sato,Tsugikazu Komoda,Kimimitsu Oda,

A convenient method for measuring immune complexes between tissue-nonspecific alkaline phosphatase (TNSALP) and immunoglobulin G (IgG) (i.e., TNSALP-IgG) would be highly useful for routine analyses. Here, we identified a surface-active agent that would dissolve membrane but not dissociate TNSALP-IgG complexes. Next, we developed an enzyme-linked immunosorbent assay (ELISA) method for detecting TNSALP-IgG complexes with two monoclonal antibodies (MoAbs): 3-29-3R was coated on assay plates and captured TNSALP-IgG from a specimen; an horseradish peroxidase (HRP)-conjugated anti-human IgG1 then reacted with captured TNSALP-IgG to form an "immunocomplex sandwich." The immunocomplex was detected via the absorbance of an HRP substrate, resulting in a semiquantitative assay. The mean absorbance of 0.195 (n=5) measured in sera from healthy donors was designated as an arbitrary unit (AU/mL) of TNSALP-IgG concentration. The ELISA values of patient sera known to contain TNSALP-IgG complexes were greater than those of normal sera (normal, 1.86 plusmn; 0.61; patient, 9.30 plusmn; 5.44), and these data were confirmed by electrophoresis. Thus, the ELISA could detect TNSALP-IgG complexes. The intraassay coefficient of variation (CV) was within 7.4% and analytical recovery was excellent. There was no significant interference from hemolytic, lipemic, or icteric serum. In summary, an ELISA using 3-29-3R MoAb and HRP-conjugated anti-human IgG1 constitutes a reliable and convenient method for the semiquantitative detection of TNSALP-IgG complexes in human serum.

Development of an ELISA method for detecting immune complexes between tissue-nonspecific alkaline phosphatase and immunoglobulin G. Publishing Authors By Initials

K Hocchi,T Ohashi,T Miura,K Sasagawa,Y Sato,F Nomura,T Tomonaga,M Sunaga,R Kojima,K Katayama,T Kato,T Sato,T Komoda,K Oda,K Hocchi,T Ohashi,T Miura,K Sasagawa,Y Sato,F Nomura,T Tomonaga,M Sunaga,R Kojima,K Katayama,T Kato,T Sato,T Komoda,K Oda,

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Development of an ELISA method for detecting immune complexes between tissue-nonspecific alkaline phosphatase and immunoglobulin G. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Journal of clinical laboratory analysis

VOLUME: 21

Page Numbers: 322-9

Journal Abbreviation: J. Clin. Lab. Anal.

ISSN: 0887-8013

DAY: 17

MONTH: 09

YEAR: 2007

Development of an ELISA method for detecting immune complexes between tissue-nonspecific alkaline phosphatase and immunoglobulin G. Information

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LANGUAGE: eng

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Grant and Affiliation Information for Development of an ELISA method for detecting immune complexes between tissue-nonspecific alkaline phosphatase and immunoglobulin G.

AFFILIATION: Division of Biochemistry, Niigata University Graduate School of Medicine and Dental Sciences, Niigata, Japan.

Country: United States

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MEDLINETA: J Clin Lab Anal

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