Development of Next-generation Peptide Affinity Tags for Enhanced Recombinant Protein Purification Using Zinc Chelation
Document Type
Thesis
Degree Name
Master of Science (MS)
Department
Chemistry
Date of Award
9-1-2026
Abstract
Metal-chelating peptide affinity tags are crucial for recombinant protein purification, but efficient purification of recombinant proteins remains a fundamental challenge in biotechnology, structural biology, and pharmaceutical research. Affinity-based purification techniques for example, Immobilized metal affinity chromatography (IMAC) combined with polyhistidine affinity tags is one of the most widely used purification methods due to its simplicity and efficiency. However, conventional His-tag purification systems often suffer from nonspecific protein binding, metal ion leakage, and reduced selectivity in complex biological mixtures. These limitations highlight the need for improved affinity tags capable of stronger and more selective metal coordination. This research aims to develop next-generation peptide affinity tags (Zn-tag) that utilize zinc chelation for enhanced recombinant protein purification. Novel histidine and cysteine containing peptide sequences such as ac-HCGPYHC, ac-CHGPYHC, and ac-CHGPYCH will be designed and analyzed to investigate their ability to coordinate Zn²⁺ ions through multidentate interactions. In this research, the metal-binding properties of these peptides will be examined and evaluated how the three Zn-tags bound and eluted from zinc resin commonly used in immobilized metal affinity chromatography using advanced analytical techniques such as UV-visible spectroscopy, high-performance liquid chromatography (HPLC), and ion mobility-mass spectrometry (IM-MS) competitive binding experiments. By systematically analyzing peptide-zinc interactions and comparing binding affinities among designed peptides, this study aims to identify optimized peptide motifs capable of serving as improved affinity tags. The results are expected to contribute to the development of mechanistic insights into Zn²⁺ driven peptide metal interactions that define binding stability, more selective, and efficient purification strategies for recombinant proteins, guiding the design of next-generation metal-chelating affinity tags with potential applications in biotechnology, proteomics, and pharmaceutical research.
Advisor
Laurence A. Angel
Subject Categories
Chemistry | Physical Sciences and Mathematics
Recommended Citation
Taleb, Md Abu, "Development of Next-generation Peptide Affinity Tags for Enhanced Recombinant Protein Purification Using Zinc Chelation" (2026). Electronic Theses & Dissertations. 1390.
https://lair.etamu.edu/etd/1390
