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Related Concept Videos

Hybridoma Technology01:31

Hybridoma Technology

Hybridoma technology is used for the large-scale production of monoclonal antibodies. Monoclonal antibodies bind to only a single antigenic determinant or epitope. Such antibodies are used in research, diagnostics, and disease therapy. The hybridoma technology established in 1975 by Georges Köhler and Cesar Milstein was awarded the Nobel Prize in Medicine in 1984 for revolutionizing research and therapy.
Hybridoma Selection
Commonly used fusion techniques — electroporation, polyethylene glycol...
Differentiation of Common Myeloid Progenitor Cells01:15

Differentiation of Common Myeloid Progenitor Cells

Common myeloid progenitors (CMPs) are oligopotent cells that can differentiate into granulocytes and macrophages. Granulocytes and macrophages are essential for protecting the body against bacterial, viral, or fungal infections. They migrate from the bone marrow into the circulating blood to reach specific tissue sites where they differentiate and help in immune surveillance. However, they survive only for a few days and must be continuously made available to the organism to maintain a robust...

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Related Experiment Video

Updated: Jun 4, 2026

Generation of Murine Monoclonal Antibodies by Hybridoma Technology
09:42

Generation of Murine Monoclonal Antibodies by Hybridoma Technology

Published on: January 2, 2017

The monoclonal myth.

Thomas C Scanlon1, Heather L Jewell, Elizabeth C Gray

  • 1Thayer School of Engineering, Dartmouth College, Hanover, NH, USA.

Bioengineered Bugs
|February 18, 2011
PubMed
Summary
This summary is machine-generated.

Multiple Vector Transformants (MVT) in yeast expression systems can lead to experimental errors. This study identifies MVT risks and provides a method to eliminate them for accurate protein engineering.

Keywords:
combinatorial library constructionhigh efficiency transformationmultiple vector transformantsprotein engineeringyeast plasmids

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Last Updated: Jun 4, 2026

Generation of Murine Monoclonal Antibodies by Hybridoma Technology
09:42

Generation of Murine Monoclonal Antibodies by Hybridoma Technology

Published on: January 2, 2017

Generation of Recombinant Human IgG Monoclonal Antibodies from Immortalized Sorted B Cells
10:32

Generation of Recombinant Human IgG Monoclonal Antibodies from Immortalized Sorted B Cells

Published on: June 5, 2015

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Yeast Genetics

Background:

  • The standard assumption in recombinant DNA technology is a "one cell-one mutant gene-one protein variant" paradigm.
  • Recent findings in Saccharomyces cerevisiae revealed high frequencies of Multiple Vector Transformants (MVT), challenging this paradigm.
  • MVT, cells with mixed plasmids, can persist across generations, impacting experimental outcomes.

Purpose of the Study:

  • To investigate the prevalence and implications of MVT in yeast expression systems.
  • To determine if MVT are specific to certain plasmid origins of replication.
  • To develop a method for identifying and mitigating MVT in protein engineering workflows.

Main Methods:

  • Transformation of yeast with recombinant plasmid libraries.
  • Culturing and analysis of yeast populations to detect MVT.
  • Development and application of an optimized sequencing procedure for MVT identification.
  • Implementation of extended liquid outgrowths and specific archiving strategies.

Main Results:

  • MVT are prevalent not only with CEN/ARS plasmids but also with high copy 2 µm plasmids in yeast.
  • MVT can cause misidentification of mutants, loss of valuable clones, and propagation of false positives.
  • An optimized sequencing method reliably identifies MVT.
  • Extended outgrowths and sequence-verified archiving effectively eliminate MVT risks.

Conclusions:

  • MVT represent a significant challenge in yeast-based protein engineering and combinatorial library screening.
  • The developed sequencing and culturing protocols effectively mitigate MVT-associated risks.
  • Ensuring monoclonal, sequence-verified genes is crucial for accurate results in protein variant studies.