Column chromatography question??

Question

Give some practical applications for column chromatography in the biotechnology industry.

Answer 1

Affinity chromatography.  I have used this for protein purification.  You attach some specific molecule to the column, say a ligand for an enzyme. Then you pass a crude mixture over the column, and only those things that bind the ligand will be retained on the column.  Very good way to purify specific proteins.

You can also attach nucleotides, like a strand of DNA to a column.  Since DNA strands like to hybridize, you can actually use such a column to isolate DNA strands.  So, lets say you know the partial sequence of a gene.  You can put that sequence on a column, isolate DNA from a cell, and use the column to purify segments of that gene from the cell.

You can use "gel filtration" chromatography, where the small beads filling the column have pores of different size, to isolate molecules based on size.  The theory is that smaller molecules will be "trapped" in the pores, and so will take a long time to elute (exit) the column, whereas big molecules will pass straight through.

You can also use ion exchange chromatography, where the column matrix has a positive or negative charge, and will bind to molecules of opposite charge (such as proteins, and nucleotides).  You can use a pH or salt gradient to elute the column, and achieve considerable separation of the molecules that bind the column.

These are just some examples; there are many others, like reverse phase chromatography.

Answer 2

Column chromatography is used to separate substances for analysis, preparation and purification. some examples for different purposes and methods:

Column chromatography is the most effective method of protein fractionation

Partition Column Chromatography of Insulin:

http://www.jbc.org/cgi/reprint/235/12/34...

Column for use in high speed liquid chromatography for micro-analysis

http://www.freepatentsonline.com/4059523...

Radial dispersion in open tubular column chromatography

http://www3.interscience.wiley.com/cgi-b...

"The K-value was measured by column chromatography using the method of Kato et al."

for full text you have to sign in:

http://www.blackwell-synergy.com/doi/abs...

General info:

http://www.answers.com/topic/column-chro...

http://www.novasep.com/chromatography_co...