Biochem Lab Affinity

Immuno-Affinity (Antigen- Antibody) Chromatography

Matrix + Antigen -----Purifies-----antibodies

Poly (U) Chromatography

Matrix + a poly-uridylic acid, synthetic poly nucleotide(Poly U) ----Purifies----mRNA

DNA Affinity Chromatography

Matrix + single stranded or double stranded bound ---Purifies--proteins (non-specific binding )

DNA Sequence Specific Chromatography

Matrix +short double-stranded DNA oligonucleotides containing a particular nucleotide sequence----purifies-
--proteins that specifically binds to a certain DNA sequence

Dye – Binding Chromatography: They mimic nucleotides such as ATP or NADH, thus they can bind to proteins.

Matrix + dyes generated from textile industry (many side chains that has confirmations similar to protein side chains &
they mimic nucleotides such as ATP or NADH ) ---Purifies -------Proteins

Gene Fusion Approaches: the gene encoding the protein of interest is infused to a second gene encoding a protein that
is easily purified by affinity chromatography. This second protein is called the tag

Matrix + the gene encoding a protein of interest is infused to a second gene encoding a protein (tag) easily purified by AC-
---Purifies -----Proteins

Metal Chelate Chromatography

Matrix + immobilized nickel (Ni) ---Purifies--- a fusion protein in which ploy histidine tag is attached. Elusion is done by introducing imidazol