Production and evaluation of monospecific antibodies for a processing-independent sequence of human progastrin.
We have produced a series of monospecific antibodies in five rabbits by immunization with a peptide corresponding to the preprogastrin sequence 76-88. Both antibody titres (0.2 to 3.0 x 10(6)), indexes of heterogeneity (approximately 1.0), and binding affinities (Koeff approximately 0.2 to 3.7 x 10(12) 1 x mol-1) were very high. The specificity for the N-terminus of the progastrin fragment was unique because even a conservative amino acid substitution five positions from the centre of binding diminished the binding by 90% for all antibodies. We conclude that these monospecific antibodies are well suited for development of a processing-independent analysis of progastrin and its products.
Bardram
L
L
Department of Gastrointestinal Surgery, Rigshospitalet, University of Copenhagen, Denmark.
Rehfeld
J F
JF
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D000941:Antigens / Q000276:immunology; D005753:Gastric Mucosa / Q000737:chemistry; D005755:Gastrins / Q000032:analysis / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007114:Immunization; D007457:Iodine Radioisotopes; D007553:Isotope Labeling; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology*; D011498:Protein Precursors / Q000737:chemistry / Q000276:immunology*; D013552:Swine
D000906:Antibodies; D000941:Antigens; D005755:Gastrins; D007106:Immune Sera; D007457:Iodine Radioisotopes; D010446:Peptide Fragments; D011498:Protein Precursors; C013706:big gastrin; C042873:preprogastrin