Synthesis and characterization of a recombinant fragment of human alpha-fetoprotein with antigenic selectivity versus albumin.
A DNA sequence coding for human alpha-fetoprotein amino acid sequence 38-119 was synthesized and cloned in a bacterial expression vector. The alpha-fetoprotein sequence was selected as the least homologous to albumin, since the two proteins have an overall amino acid identity of approximately 38%. A chimeric protein was obtained which was purified by preparative electrophoresis and characterized in its primary structure by fast atom bombardment mass spectometry. About 70% of the alpha-fetoprotein sequence was physically mapped and found to correspond to the amino acids encoded in the synthetic gene. The use of this recombinant protein allowed the selection of monoclonal antibodies recognizing both the recombinant fragment and native alpha-fetoprotein. These antibodies should allow the development of an immunoassay for alpha-fetoprotein with absolute selectivity versus albumin. This might result in more sensitive clinical determinations, avoiding the possibility of cross-reactions.
Giuliani
M M
MM
Cell and Molecular Biology Laboratories, Sclavo Research Center, Siena, Italy.
Ricci
S
S
Ratti
G
G
Pucci
P
P
Marino
G
G
Malorni
A
A
Ceccarini
C
C
Terrana
B
B
Tecce
M F
MF
D000418:Albumins / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003429:Cross Reactions; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D013058:Mass Spectrometry; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D000509:alpha-Fetoproteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology
D000418:Albumins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D000509:alpha-Fetoproteins