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Protein purification

Conference paper
Part of the Advances in Biochemical Engineering/Biotechnology book series (ABE, volume 40)

Abstract

This monograph summarizes recent developments in the purification and analysis of natural and recombinant proteins. The basic strategies employed in protein purification are reviewed with regards to the characteristics of the protein of interest that may aid its isolation, choice of the starting material, and use of denaturants. Preparation of cell-free extracts followed by bulk precipitation and/or phase partition constitute the initial steps of many purification schemes. Chromatographic methods (size exclusion, ion exchange, hydroxylapatite, reversed phase, hydrophobic interaction and affinity based) utilizing either traditional, low pressure or high-performance liquid chromatography instrumentation are discussed. Electrophoretic techniques used to analyze the homogeneity of the protein product include SDS-PAGE, isotachophoresis, IEF and two dimensional gel electrophoresis.

Keywords

Sodium Dodecyl Sulfate Size Exclusion Chromatography Protein Purification Purification Scheme Hydrophobic Interaction Chromatography 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations

a

selectivity factor

ABx

antibody exchanger

cDNA

complementary DNA

CM

carboxymethyl

CoA

coenzyme A

D

diameter

Da

Daltons

DEAE

diethylaminoethyl

DNA

deoxyribonucleic acid

FPLC

fast protein liquid chromatography

FR

flow rate

HETP

height equivalent to a theoretical plate

HIV-1

human immunodeficiency virus type 1

HPLC

high-performance liquid chromatography

IEF

isoelectric focusing

IgG

immunoglobulin G

IgM

immunoglobulin M

K

distribution coefficient

k′

capacity factor

L

length

LV

linear velocity

N

number of theoretical plates

PAGE

polyacrylamide gel electrophoresis

PEI

polyethyleneimine

PEG

polyethylene glycol

pI

isoelectric point

PMSF

phenylmethylsulfonyl fluoride

QAE

quaternary aminoethyl

R

resolution

RNA

ribonucleic acid

Rt

retention time

SDS

sodium dodecyl sulfate

SF

scale-up factor

SP

sulfoxypropyl

TFA

trifluoroacetic acid

V

retention volume

V0

void volume

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Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  1. 1.Biotech Research Laboratories, Inc.RockvilleUSA
  2. 2.Philip Morris Research CenterRichmondUSA

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