Enzyme immunoassay of (β-hexosaminidase isoenzymes using monoclonal antibodies

Abstract

Purified β-hexosaminidase A (Hex A) and β-hexosaminidase B (Hex B) were used as immunogens in mice, with the purpose of obtaining isoenzyme-specific monoclonal antibodies (mabs). A total of 60 mabs were developed, 23 specific for Hex A and 37 recognizing both isoenzymes. At low pH, two of the latter mabs reacted only with Hex B, and it was, therefore, possible to develop enzyme immunoassays for the specific determination of Hex A and Hex B. The precision of the methods was adequate with intra- and interassay coefficients of variation below 3%.

Key Words:

• β-hexosaminidase
• EIA
• isoenzymes
• lysosomal storage diseases
• monoclonal antibodies
• renal diseases