SIMULTANEOUS QUANTITATION OF SPECIFIC IGE AGAINST 20 PURIFIED ALLERGENS IN ALLERGIC PATIENTS SERA BY CHECKERBOARD IMMUNOBLOTTING (CBIB)

A simple technique, checkerboard immunoblotting (CBIB), is described f or simultaneous quantitation of specific IgE antibodies against severa l allergens in human sera. Using as little as 50 mu l of each of the 2 0 sera examined against 20 different allergens, it was possible in a s ingle run to achieve 400 tests. To guarantee high specificity and sens itivity of the assay, this new application of CBIB employs purified al lergens, cyanogen bromide-activated nitrocellulose membrane, and Phosp horimager technology. Results are expressed both qualitatively (five c lasses) and quantitatively in kilo units per liter equilibrated agains t the World Health Organization (WHO) standard for IgE. There was exce llent agreement between the results of CBIB and the results of Pharmac ia Cap System, an alternative method widely used for measuring serum-s pecific IgE. The CBIB method certainly could be useful in any laborato ry interested in allergy clinical research for easy screening and rela tive quantitation of allergen-specific human IgE. (C) 1997 Wiley-Liss, Inc.