Negative chromatography: Progress,applications and future perspectives |
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Affiliation: | 1. Chemical Engineering Discipline, School of Engineering, Monash University Malaysia, Jalan Lagoon Selatan, Bandar Sunway, Selangor 46150, Malaysia;2. Multidisciplinary Platform of Advanced Engineering, Monash University Malaysia, Jalan Lagoon Selatan, Bandar Sunway, Selangor 46150, Malaysia;1. The University of Tokyo, Tokyo, Japan;2. Max-Planck-Institut für Plasmaphysik, EURATOM Association, Garching, Germany;1. Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China;2. School of Biological Science and Biotechnology, Minnan Normal University, Zhangzhou 363000, China |
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Abstract: | In negative chromatography, the impurities bind on the adsorbent, and the product is allowed to flow through the chromatographic column. Negative chromatography is an alternative to positive chromatography under certain circumstances and has been used to purify various biomolecules. For this review, a detailed survey of the performance of reported studies on negative chromatography was conducted. The applications of negative chromatography in the capture and intermediate purification steps for biomolecules (e.g., plasmid DNA, antibodies, enzymes, hemoglobin, virus particles and cells) are reviewed. The negative chromatographic adsorbents adsorb the impurities through surface charge, hydrophobic interaction at specific sites on the surface, hydrophobic interaction, hydrogen bonding and functional groups. Examples of applications of negative chromatography according to the type of chromatography matrix used are summarized and discussed. In addition, the effects of operating conditions (initial protein concentration, buffer ions, pH and salt concentration) are discussed, and the criteria for choosing negative or positive chromatography are summarized. The literature survey showed that there will be future limitations and challenges ahead in implementation of negative chromatography. Possible solutions to the limitations and challenges of negative chromatography and future trends for developing negative chromatography are discussed. |
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Keywords: | Negative chromatography Flow-through chromatography Protein purification Adsorbent Recombinant protein Downstream bottleneck |
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