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Figure 3 | BMC Biotechnology

Figure 3

From: An improved phage-display panning method to produce an HM-1 killer toxin anti-idiotypic antibody

Figure 3

Identification of positive clones to immobilized antigen. The binding properties of randomly selected clones to nmAb-KT were measured by microtiter plate ELISA. The supernatant containing soluble scFv antibodies was added to each well of a microtiter plate that had been pre-coated with nmAb-KT and detection was done by HRP-conjugated anti-E-tag antibody. A. Regular panning: 12 of 50 (24%) tested clones of infected E. coli TG1 contained the anti-idiotypic scFv gene of HM-1 killer toxin, and so soluble scFvs showed positive ELISA signals after the fifth round of panning. B. Modified subtractive panning: 40 of 50 (80%) tested clones showed positive ELISA signals after the fifth round of modified subtractive panning. Negative controls consisted of microtiter plates coated with blocking buffer (N-1) or washing buffer (N-2). The results were means of duplicates.

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