Synthesis of Human
Proinsulin C-peptide and Preparation of Specific Antibody to It
LI Mo-Yi, LIU Zhi-Hui, SHAO Xiao-Xia, XIA Qi-Chang, ZHOU
Guo-Ming, LIN Qi-Shui1, CUI Da-Fu*
( Institute of Biochemistry and Cell Biology, Shanghai Institutes for
Biological Sciences, the Chinese Academy of Sciences, Shanghai 200031, China； 1State Key Laboratory of Molecular
Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for
Biological Sciences, the Chinese Academy of Sciences, Shanghai 200031, China )
Abstract A HPLC and
CE pure human proinsulin C-peptide was synthesized by solid-phase method and
TSK column purification. Its amino acid sequence and MS were consistent with
theoretical values. In comparison with the formly reported chemical synthesis
of C-peptide, this method has the advantage of simplicity and higher overall
yield (41%). To improve the immunogenicity and specificity of oligopeptide
antibody, the acrylyl-C-peptides were transformed into a polymer；the product had a
poly-propionyl-core matrix with C-peptide branches. This treatment gave a
macromolecule with a Mr about 25 kD. By using the polymer
to immunize New Zealand rabbits for 30 days, specific antiserum was obtained
with titer of 2.5×104 (by ELISA), which did not cross react with
BSA. Thus, the poly-propionyl-peptide system provided a new approach for
preparing synthetic peptide antibody and therefore is promising for the
preparation of synthetic peptide-based vaccine.
Key words human C-peptide；peptide synthesis；oligo-peptide antigen
*Corresponding author：Tel,86-21-64374430-262；Fax,86-21-64338357；e-mail, [email protected]