Production of Monoclonal Antibodies Specific to Eyestalk Neuropeptides of Penaeus monodon sing Sinus Gland Section and Immunosuppression Technique

Nanthika Panchan^a,b, Paisarn Sitigorngul^c*, Parin Chaivisuthangkura^c, Siwaporn Longyant^c,Weerawan Sithigorngul^c and Amorn Petsom^a,b

ABSTRACT: Immunosuppression method was utilized for immunization in order to generate monoclonal antibodies specific to neuropeptides in the eyestalk of Penaeus monodon. Selection of peptide specific monoclonal antibodies was done by immunohistochemistry against eyestalk sections. Four monoclonal antibodies bound to sinus gland and neurons in the eyestalk were isolated. All of these antibodies bound to putative peptides in different fractions of eyestalk extract separated by RP-HPLC. The antibodies were used to monitor the presence of the peptides during purification of sinus gland extract by RP-HPLC using dot-ELISA method. Crustacean hyperglycemic hormones (Sgp I, III and IV) were isolated and identified with SG 24, 26 and 293 monoclonal antibodies respectively. An unknown peptide with molecular mass of 9127.56 daltons was identified with SG 782 antibody. The N-terminal sequencing of the first 20 residues revealed that the sequence of this peptide is a novel peptide sharing no sequence identity to any known crustacean or vertebrate.

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a Biotechnology Program, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.
b Institute of Biotechnology and Genetic Engineering, Chulalongkorn University, Bangkok 10330, Thailand.
c Department of Biology, Faculty of Science, Srinakharinwirot University, Bangkok 10110, Thailand.

* Corresponding author, E-mail: paisarn@swu.ac.th

Received 20 Aug 2004, Accepted 3 Nov 2004