BC367PS4-1

BC367PS4-1 - Brian Blanchard BC367 Biochemistry of the Cell I PS#4 Q1 Hydroxyapetite chromatography is a method of protein purification that

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BC367 Biochemistry of the Cell I PS#4 Q1 Hydroxyapetite chromatography is a method of protein purification that harnesses the affinity of negatively and positively charged regions within a residue for opposite charges on the stationary resin of HA. HA is essentially a resin in the form of a crystalline matrix decorated with negatively charged phosphate groups and positively charged calcium ions. This method of protein purification has been used extensively for antibody purification. With the correct buffer selection, this process is very effective at separating antibodies with neutral, acidic, or basic natures. For example, HA has a very high tollerence for sodium chloride as a loading medium for acidic antibodies (allows the antibodies to remain in solution), and allows for the dissociation of ionic interactions between DNA contaminants and the desired antibodies. This method had also proven useful for binding weakly interacting basic proteins by the inclusion of 1 mM phosphate into the buffer, thereby suppressing the ability of the HA’s calcium cations to repel amines. (Gagnon, P; Frost, R; Tunón, P; Ogawa, T. 2009) Gagnon, P; Frost, R; Tunón, P; Ogawa, T . CHT Ceramic Hydroxyapetite – A New Dimesion in Chromatography of Biological Molecules . Bio-Rad Tech Note 2156 (Chromatography). Bio-Rad Laboratories, Inc., Life Science Group. 2009. http://www.bio-rad.com/LifeScience/pdf/Bulletin_2156.pdf
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This note was uploaded on 10/06/2010 for the course ES ES271 taught by Professor Machaut during the Spring '10 term at Colby.

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BC367PS4-1 - Brian Blanchard BC367 Biochemistry of the Cell I PS#4 Q1 Hydroxyapetite chromatography is a method of protein purification that

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