Development of immunoenzymatic reactions of peptides using recombinant hybrid proteins.

A recombinant hybrid protein comprising the bacteriophage fr coat protein, a short linker, and atrial natriuretic factor (ANF) as well as native and recombinant phage coat proteins, ANF, and variants of the hybrid recombinant protein were used in the development of various immunological reactions including immunization, preparation of affinity columns, purification of antibodies, synthesis of conjugates, and immunoenzyme assay of ANF and the recombinant protein. The hybrid protein is effective in competitive assay of ANF and other constructs that include the phage protein.

Synthesis of recombinant atrial natriuretic peptide (rANP) using hybrid fusion protein-phage fr coat/ANP (CP/ANP).

Recombinant atrial natriuretic peptide (rANP) was expressed in and isolated from E. coli. rANP was purified using HPLC.

High level expression of alpha-human atrial natriuretic factor as a fusion polypeptide with phage fr coat protein in Escherichia coli.

A synthetic DNA sequence coding for the 28 amino acid residues of alpha-human atrial natriuretic factor (alpha-hANF) and the N-terminal linker tripeptide Ile-Asp-Lys was inserted in the 3'-terminal part of the RNA bacteriophage fr coat protein gene. The cloned hybrid gene was isolated and placed into an expression vector under the control of the inducible E. coli tryptophan promoter and phage fr coat protein translation initiation region (TIR) sequence.