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

Assessing genetic stability at the nucleic acid level

F Galibert1

  • 1Laboratoire de Recombinaisons Génétiques, Hôpital Saint-Louis-Centre Hayem, Paris, France.

Developments in Biological Standardization
|January 1, 1994
PubMed
Summary
This summary is machine-generated.

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

[Anti-COVID-19 RNA vaccines].

Bulletin de l'Academie nationale de medecine·2021
Same author

COVID-19 epidemic phases: Criteria, challenges and issues for the future.

Bulletin de l'Academie nationale de medecine·2020
Same author

[Covid-19 epidemic phases: Criteria, challenges and issues for the future].

Bulletin de l'Academie nationale de medecine·2020
Same author

Contribution of radiation hybrids to genome mapping in domestic animals.

Cytogenetic and genome research·2009
Same author

Comparative genomic mapping of uncharacterized canine retinal ESTs to identify novel candidate genes for hereditary retinal disorders.

Molecular vision·2009
Same author

The dog genome.

Genome dynamics·2008

Recombinant proteins can be contaminated by altered molecules due to genetic instability or errors during protein production. Detecting low-level contaminants requires methods beyond direct gene sequencing, such as SSCP or CCM.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Protein Engineering

Background:

  • Recombinant proteins, used as therapeutics, can be contaminated by mutated or altered molecules.
  • Genetic instability and errors in protein synthesis or modification contribute to these contaminants.
  • While protein-level alterations are not amplified, understanding genetic origins is crucial for interpretation.

Purpose of the Study:

  • To highlight the challenges in detecting low-level mutated molecules in recombinant protein production.
  • To emphasize the importance of considering genetic factors when analyzing protein products.
  • To recommend advanced analytical methods for comprehensive contaminant detection.

Main Methods:

  • Direct gene sequencing of plasmids or RT-PCR products.

Related Experiment Videos

  • Analysis of molecular alterations at the nucleic acid and protein levels.
  • Application of alternative methods like Single-Strand Conformational Polymorphism (SSCP) and Chemical Cleavage Modification (CCM).

    • Main Results:

    • Direct sequencing methods can miss low-level mutated contaminants.
    • Nucleic acid alterations are amplified, affecting a larger proportion of molecules compared to protein-level changes.
    • Advanced techniques are necessary for accurate detection and characterization of contaminants.

    Conclusions:

    • Accurate assessment of recombinant protein purity requires understanding genetic system variations.
    • Statistical analysis of population polymorphism and methods like SSCP and CCM are recommended for detecting low-level contaminants.
    • Robust analytical strategies are essential for ensuring the safety and efficacy of protein-based therapeutics.