Chromogenic Immuno detection of Human Serum Albumin and α-l-Fucosidase Clones in a Human Hepatoma cDNA Expression Library

Abstract

A human hepatoma cDNA library was constructed in λgt11, a bacteriophage vector that was designed to express cDN A-encoded antigenic determinants in Escherichia coli. The cDNA expression library contained ∼ 8 × 10⁶ recombinant phages with an average insert size of 780 bp. The feasibility of using a chromogenic immunodetection procedure to isolate cDNA clones was proved by isolating a human serum albumin (HSA) cDNA clone. An ∼ 1.0-kb cDNA clone was then isolated by screening the library with rabbit anti-human α-l-fucosidase antibodies. The identity of the α-l-fucosidase cDNA clone was confirmed by DNA sequence analysis and a comparison of the predicted amino acid sequence to the amino acid sequences of human α-l-fucosidase tryptic peptides.