wet chemical precipitation method by JAGJIT SINGH KAHLON

1. CHEMICAL SYNTHESIS OF HYDROXYAPATITE FOR APPLICATION IN CHROMATOGRAPHY BY Jagjit Singh Kahlon Sec- F77E1 Roll No. 49 Faculty Adviser: Anjuvan Singh 15097 Biotechnology LSSP Department of Biotechnology Lovely Institute of Engineering and Computer Applications (LIECA) LPU, Phagwara, India

2. CONTENTS INTRODUCTION OBJECTIVE REVIEW LITERATURE RESEARCH METHODOLOGY RESULT

6. Literature Review Protein chromatography on hydroxyapatite columns. Cummings LJ , Snyder MA , Brisack K . Bio-Rad Laboratories, Inc., Hercules, California, USA. Abstract The introduction of spherical forms of hydroxyapatite has enabled protein scientists to separate and purify proteins multiple times with the same packed column. Biopharmaceutical companies have driven single column applications of complex samples to simpler samples obtained from upstream column purification steps on affinity, ion exchange or hydrophobic interaction columns. Multiple column purification permits higher protein loads to spherical forms of hydroxyapatite and improved reduction in host cell protein, aggregates, endotoxin, and DNA from recombinant proteins. The chemical interactions of hydroxyapatite surface with common ions, metals, and phosphate species affect column lifetimes. Column packing methods for the robust spherical particles as well as the microcrystalline hydroxyapatite particles are reviewed. Applications covering extracted proteins and recombinant protein purification, especially monoclonal antibodies, with multiple chromatography media in concert with hydroxyapatite are reviewed.

7. Literature Review Purification of lactoferrin using hydroxyapatite. Ng PK , Yoshitake T . Process Applications R&D, Process Chromatography Division, Bio-Rad Laboratories, Hercules, CA 94547, USA. paul ng@bio-rad.com Abstract Lactoferrin is an important nutriceutical with various physiological functions. It is present in whey at very low concentrations. This work describes a mixed-mode (hydroxyapatite) chromatography method for one-column fractionation of lactoferrin from whey. Lactoperoxidase, a protein with similar molecular weight and isoelectric point, was initially desorbed from the matrix under isocratic conditions. Lactoferrin was obtained in homogeneity without lactoperoxidase activity and free from other major whey proteins such as alpha lactoalbumin and beta lactoglobulin.

10. Research Methodology

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