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

A carbohydrate binding module as a diversity-carrying scaffold.

L Cicortas Gunnarsson1, E Nordberg Karlsson, A-S Albrekt

  • 1Department of Immunotechnology, Lund University, PO Box 7031, S-220 07 Lund, Sweden.

Protein Engineering, Design & Selection : PEDS
|April 15, 2004
PubMed
Summary

Researchers developed novel binding proteins using a stable scaffold, CBM4-2, for biotechnology. This engineered scaffold demonstrated high productivity and stability, enabling diverse applications.

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

Warming weather changes the chemical composition of oat hulls.

Plant biology (Stuttgart, Germany)·2021
Same author

Porcine field fertility with two different insemination doses and the effect of sperm morphology.

Reproduction in domestic animals = Zuchthygiene·2006
Same author

No signs of immunoactivation in the cerebrospinal fluid during treatment with infliximab.

Annals of the rheumatic diseases·2006
Same author

Pregnancy rates in lactating Holstein-Friesian cows after artificial insemination with sexed sperm.

Reproduction in domestic animals = Zuchthygiene·2006
Same author

Transplantation of normal boar testicular cells resulted in complete focal spermatogenesis in a boar affected by the immotile short-tail sperm defect.

Reproduction in domestic animals = Zuchthygiene·2006
Same author

Inverse relationship between central serotonergic neurotransmission and blood pressure in alcohol-dependent male subjects.

Journal of neural transmission (Vienna, Austria : 1996)·2006

Area of Science:

  • Biotechnology
  • Protein Engineering
  • Molecular Biology

Background:

  • The biotechnology sector requires binding proteins with enhanced specificity, affinity, stability, and productivity for large-scale applications.
  • Naturally occurring binding proteins are often diversified to create libraries for discovering molecules with novel properties, serving as scaffolds.

Purpose of the Study:

  • To evaluate the thermostable carbohydrate-binding module CBM4-2 from Rhodothermus marinus as a scaffold for generating binding proteins with diverse specificities.
  • To create and screen a combinatorial library based on CBM4-2 by introducing variations in its carbohydrate-binding site.

Main Methods:

  • Construction of a combinatorial library by diversifying 12 positions in the CBM4-2 carbohydrate-binding site.
  • Selection of variants using phage display against carbohydrate polymers (birchwood xylan, Avicel, ivory nut mannan) and a glycoprotein (human IgG4).

Related Experiment Videos

  • Assessment of protein productivity in Escherichia coli and measurement of thermal stability (melting transition temperature).

    • Main Results:

    • Successful selection of CBM4-2 variants with specificity towards different carbohydrate polymers and human IgG4 from a library of 1.6 x 10^6 clones.
    • High average productivity of purified protein (69 mg/l culture) when expressed in Escherichia coli.
    • All selected variants were stable, exhibiting a high melting transition temperature of 75.7 ± 5.3 °C.

    Conclusions:

    • The CBM4-2 molecule is a highly suitable and versatile scaffold for developing novel binding proteins.
    • Engineered variants derived from CBM4-2 demonstrate potential for various biotechnological applications due to their stability and productivity.