Substitution of carbonate buffer by water for IgG immobilization in enzyme linked immunosorbent assay

Journal of Immunoassay & Immunochemistry
Tulsidas G ShrivastavKiran P Kariya

Abstract

The first step of enzyme linked immunosorbent assay (ELISA), namely, adsorption of antigen or antibody to the plastic microtiter well plate, was studied as a function of insolubility of IgG in water. Immobilization efficiency was assessed in terms of number of wells coated per milliliter of primary antiserum. We have compared different coating/immobilization protocols, i.e., direct and indirect immobilization of primary antibody to the plastic microtiter well plate using carbonate buffer and phosphate buffer with glutaraldehyde. We have observed efficient coating when the immobilization of primary antibody through an immunobridge technique was performed, where water was used as a coating medium. It gave a higher number of wells coated per milliliter of anti-serum (primary or secondary) than other compared coating protocols and it allowed the use of serum (non-immune) and anti-serum (primary and secondary antibody) dilutions, avoiding the need for gamma-globulin purification from normal and immunized serum.

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Citations

Oct 6, 2004·Journal of Immunoassay & Immunochemistry·Tulsidas G Shrivastav
Feb 22, 2014·Monoclonal Antibodies in Immunodiagnosis and Immunotherapy·Bin WangShi-quan Han
Mar 19, 2014·Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy·Songcheng YuYongjun Wu
Mar 25, 2008·Journal of Immunoassay & Immunochemistry·V TripathiTulsidas G Shrivastav
Dec 13, 2005·Clinica Chimica Acta; International Journal of Clinical Chemistry·Anupam BasuTulsidas G Shrivastav
Jul 7, 2011·Journal of Immunoassay & Immunochemistry·Tulsidas G ShrivastavBhavana Pandey
Jul 7, 2011·Journal of Immunoassay & Immunochemistry·Tulsidas G ShrivastavRita Singh
Mar 25, 2014·Journal of Immunoassay & Immunochemistry·Tulsidas G ShrivastavDinesh Kumar
Sep 13, 2005·Journal of Immunoassay & Immunochemistry·Anupam BasuSaumen Kumar Maitra
Mar 31, 2009·Journal of Immunoassay & Immunochemistry·Sabana KhatunTulsidas G Shrivastav
Dec 18, 2007·Journal of Immunoassay & Immunochemistry·S NaraH Singh
Sep 30, 2008·Journal of Immunoassay & Immunochemistry·Seema NaraTulsidas G Shrivastav
Apr 6, 2007·The Journal of Veterinary Medical Science·Shun-ichi NagataMasao Kuwajima
May 17, 2011·Journal of Immunoassay & Immunochemistry·Tulsidas G ShrivastavRita Singh
Nov 30, 2010·Journal of Immunoassay & Immunochemistry·Tulsidas G ShrivastavDinesh Kumar
Dec 21, 2011·Journal of Immunoassay & Immunochemistry·Tulsidas G ShrivastavBhavana Deshmukh
Jun 29, 2012·Journal of Immunoassay & Immunochemistry·Tulsidas G ShrivastavDinesh Kumar
Nov 30, 2010·Journal of Immunoassay & Immunochemistry·Tulsidas G ShrivastavAnjali Nagendra
Jan 18, 2013·Journal of Immunoassay & Immunochemistry·Tulsidas G ShrivastavDinesh Kumar

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