Journal of Pure and Applied MicrobiologyVol. 9 No. 3

Identification of Mammary Tumour Homing Peptides by In vivo Biopanning using Phage Display Library

Harish D. Ramachandra1*, Arvind K. Singh1, Shishir K. Gupta1, Madhu C. Lingaraju2, Goravanahalli M. Vidyalakshmi3, Aditya P. Sahoo1, G. Ravikumar1 and Ashok K. Tiwari4

1Division of Veterinary Biotechnology, 2Division of Pharmacology and Toxicology, 3Division of Physiology and Climatology, 4Division of Biological Standardization, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, U.P., 243 122, India.

Received on 11 May 2015 and accepted on 15 June 2015

 

ABSTRACT

Mammary tumour was induced by subcutaneous injections of LA7 cells (approximately 106 cells) into the mammary gland pad of 50 day old female Sprague-Dawley rats. Mammary tumour was observed at the site of injection of cells in sixteen out of thirty rats. The tumour was confirmed by physical examination and histopathology. We conducted four rounds of in vivo biopanning using M13 phage display library (7-mer) in mammary tumour bearing rats. The tumour binding phages were rescued and amplified by E coli ER2738. Then, sixty plaque-forming units (pfus) were selected randomly, each phage clone was amplified and DNA from individual clone was isolated. Afterwards pIII gene which contains the inserted peptide coding DNA sequence was confirmed by PCR and DNA sequencing. These sequences were analyzed by MegAlign (DNASTAR) software and identified consensus peptide motif ‘SPPR’. This motif was predicted that, it is having mammary tumour homing ability and maybe useful for tumour targeted delivery of therapeutics and imaging agents.

Keywords : Mammary tumour, LA7 cells, In vivo biopanning, Homing peptides, Motifs.