Treatment of human muscle creatine kinase with glutaraldehyde preferentially increases the immunogenicity of the native conformation and permits production of high-affinity monoclonal antibodies which recognize two distinct surface epitopes.

Journal of Immunological Methods
N T ManG E Morris

Abstract

Treatment of human muscle creatine kinase (MM-CK) with glutaraldehyde produced highly aggregated forms which retained the native antigenic structure. Immunization of BALB/c mice with CK aggregates instead of untreated CK produced over ten-fold higher titres of antibody against native CK without increasing the titres of antibody against denatured enzyme. Production of high-affinity monoclonal antibodies specific for both the muscle isoenzyme and the native conformation became possible where the use of untreated CK had failed. Four monoclonal antibodies have been characterized by an epitope mapping technique and compared with a commercially available monoclonal antibody. One antibody has a much higher affinity for MM-CK than the other three and the commercial antibody. Competition studies show that it also recognizes a different epitope on the CK surface from the other three monoclonal antibodies which bind to the same surface region as the commercial antibody. Immunoassays based on the high affinity antibody can easily measure less than 1 ng of CK, a sensitivity comparable to, or better than, standard enzymatic assays.

References

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Jan 1, 1981·Methods in Enzymology·M J O'Sullivan, V Marks

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Citations

Feb 5, 2003·Toxicon : Official Journal of the International Society on Toxinology·Long-sen ChangChun-chang Chang
Jan 30, 1995·FEBS Letters·T M NguyenG E Morris
May 2, 2012·International Journal of Pharmaceutics·Wei WangSandeep Nema
Sep 5, 2009·Journal of Virological Methods·Haggag S ZeinKazutaka Miyatake
Jun 17, 2005·Journal of Pharmaceutical Sciences·Angus M Sinclair, Steve Elliott
Feb 4, 2006·The Journal of Immunology : Official Journal of the American Association of Immunologists·Paula M BerguerFernando A Goldbaum
Jul 6, 1990·Biochimica Et Biophysica Acta·G E Morris, A J Cartwright

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