Two-stage screening of combinatorial peptide libraries. Application to bovine serum albumin ligand selection

Authors

  • María Camila Martínez-Ceron NANOBIOTEC Institute. UBA-CONICET. Cathedra of Biotechnology. School of Pharmacy and Biochemistry. National Scientific and Technological Research Council (CONICET).
  • Silvana Laura Giudicessi NANOBIOTEC Institute. UBA-CONICET. Cathedra of Biotechnology. School of Pharmacy and Biochemistry. National Scientific and Technological Research Council (CONICET).
  • Johanna Nataly Kruszyn NANOBIOTEC Institute. UBA-CONICET. Cathedra of Biotechnology. School of Pharmacy and Biochemistry.
  • Mariela Mirta Marani National Scientific and Technological Research Council (CONICET). National Patagonian Center (CENPAT)-CONICET.
  • Fernando Albericio Institute for Research in Biomedicine. Department of Organic Chemistry, University of Barcelona. CIBER-BBN, Networking Centre on Bioengineering, Biomaterials and Nanomedicine School of Chemistry, Yachay Tech, Yachay City of Knowledge
  • Osvaldo Cascone NANOBIOTEC Institute. UBA-CONICET. Cathedra of Biotechnology. School of Pharmacy and Biochemistry. National Scientific and Technological Research Council (CONICET).
  • Silvia Andrea Camperi NANOBIOTEC Institute. UBA-CONICET. Cathedra of Biotechnology. School of Pharmacy and Biochemistry. National Scientific and Technological Research Council (CONICET).

Keywords:

combinatorial libraries, peptides, affinity, solid phase

Abstract

Short peptides are excellent ligands for affinity chromatrography. Divide-couple-recombine method allows obtaining a peptide library with all possible combinations of the amino acids in the form of "one bead-one peptide". It was designed a two-stage library screening method for peptide ligands selection in this work. The library screening was performed with the target protein (bovine seroalbumin, BSA) coupled to Texas Red. Fluorescent beads were automatically isolated using the Complex Object Parametric Analyzer and Sorter (COPAS) equipment. Isolated beads were washed. The second screening was performed with BSA coupled to biotin and with streptavidin-peroxidase and revealed with 3,3'-diaminobenzidine. Peptides from isolated beads were sequenced using a mass spectrometry analyzer. Those peptides appearing with more frequency were synthesized and immobilized on agarose. All peptides adsorbed BSA but not Texas-Red nor streptavidin-peroxidase. As BSA from fetal bovine serum is the main contaminant, when expressing rhEPO in CHO cell culture, it was studied rhEPO adsorption on these matrices for their future application in BSA removal from cell cultures. When a pure sample of rhEPO was loaded on peptide-agarose columns, all rhEPO passed through.

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Published

2018-04-24

How to Cite

Martínez-Ceron, M. C., Laura Giudicessi, S., Nataly Kruszyn, J., Mirta Marani, M., Albericio, F., Cascone, O., & Andrea Camperi, S. (2018). Two-stage screening of combinatorial peptide libraries. Application to bovine serum albumin ligand selection. NATIONAL CENTER FOR SCIENTIFIC RESEARCH (CENIC) BIOLOGICAL SCIENCES JOURNAL, 46(1), 77-86. Retrieved from https://revista.cnic.cu/index.php/RevBiol/article/view/96

Issue

Section

Research articles