Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Affinity precipitation of proteins

M N Gupta1, R Kaul, D Guoqiang

  • 1Department of Biotechnology, Lund University, Sweden.

Journal of Molecular Recognition : JMR
|September 1, 1996
PubMed
Summary

Affinity precipitation uses special ligands to selectively isolate proteins early in downstream processing. This method controls protein separation by altering polymer solubility with pH or temperature.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Medical implications of protein moonlighting.

The Indian journal of medical research·2019
Same author

Nanoenvironmental Effects Dramatically Influence the Sensitivity of Immunoassays.

Trends in biotechnology·2017
Same author

Retrospective analysis of spinal trauma in patients with ankylosing spondylitis: a descriptive study in Indian population.

Spinal cord·2014
Same author

Structure and functional investigation of ligand binding by a family 35 carbohydrate binding module (CtCBM35) of β-mannanase of family 26 glycoside hydrolase from Clostridium thermocellum.

Biochemistry. Biokhimiia·2014
Same author

Chemically modified poly(2-hydroxyethyl methacrylate) cryogel for the adsorption of heparin.

Journal of biomedical materials research. Part B, Applied biomaterials·2014
Same author

Thermus 2S from Thai hot springs: isolation and immobilization.

World journal of microbiology & biotechnology·2014

Area of Science:

  • Biotechnology
  • Protein Chemistry
  • Separation Science

Background:

  • Downstream processing of proteins requires efficient isolation techniques.
  • Selective protein isolation is crucial for various biotechnological applications.
  • Current methods may lack efficiency or selectivity in early stages.

Purpose of the Study:

  • To investigate affinity precipitation as an early-stage downstream processing technique.
  • To demonstrate the selective isolation of proteins using heterobifunctional ligands.
  • To explore the integration of affinity precipitation with other separation methods.

Main Methods:

  • Utilizing heterobifunctional ligands with specific protein affinity and controllable polymer solubility.
  • Inducing precipitation by altering polymer solubility via pH or temperature.
  • Employing natural polymers (chitosan, alginate) and synthetic polymers (Eudragit S-100, polyethyleneimine).
  • Coupling aromatic dyes (Cibacron Blue 3GA) to polymers for enzyme affinity precipitation.
  • Integrating affinity precipitation with aqueous two-phase systems for cell debris removal.

Main Results:

  • Successful selective isolation of target proteins using affinity precipitation.
  • Demonstrated reversible solubility control of polymers by adjusting pH or temperature.
  • Chitosan and alginate effectively isolated wheat germ agglutinin and pectinase.
  • Cibacron Blue 3GA coupled to polymers enabled affinity precipitation of model enzymes like lactate dehydrogenase and alcohol dehydrogenase.
  • Integration with aqueous two-phase systems showed promise for combined extraction and purification.

Conclusions:

  • Affinity precipitation is a viable technique for early-stage selective protein isolation.
  • Heterobifunctional ligands offer tunable control over protein precipitation.
  • The method is adaptable using various natural and synthetic polymers.
  • Potential for integration with other separation techniques enhances its applicability.

Related Experiment Videos