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HybriFree: a robust and rapid method for the development of monoclonal antibodies from different host species

Overview of attention for article published in BMC Biotechnology, January 2016
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  • Good Attention Score compared to outputs of the same age (76th percentile)
  • Average Attention Score compared to outputs of the same age and source

Mentioned by

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3 tweeters
patent
1 patent

Citations

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10 Dimensions

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71 Mendeley
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Title
HybriFree: a robust and rapid method for the development of monoclonal antibodies from different host species
Published in
BMC Biotechnology, January 2016
DOI 10.1186/s12896-016-0232-6
Pubmed ID
Authors

Gaily Kivi, Kaupo Teesalu, Jüri Parik, Elen Kontkar, Mart Ustav, Liis Noodla, Mart Ustav, Andres Männik

Abstract

The production of recombinant monoclonal antibodies in mammalian cell culture is of high priority in research and medical fields. A critical step in this process is the isolation of the antigen-binding domain sequences of antibodies possessing the desired properties. Many different techniques have been described to achieve this goal, but all have shortcomings; most techniques have problems with robustness, are time-consuming and costly, or have complications in the transfer from isolation to production phase. Here, we report a novel HybriFree technology for the development of monoclonal antibodies from different species that is robust, rapid, inexpensive and flexible and can be used for the subsequent production of antibodies in mammalian cell factories. HybriFree technology is illustrated herein via detailed examples of isolating mouse, rabbit and chicken monoclonal antibody sequences from immunized animals. Starting from crude spleen samples, antigen capturing of specific B-cells is performed initially. cDNA of antibody variable domains is amplified from the captured cells and used a source material for simple and rapid restriction/ligation free cloning of expression vector library in order to produce scFv-Fc or intact IgG antibodies. The vectors can be directly used for screening purposes as well as for the subsequent production of the developed monoclonal antibodies in mammalian cell culture. The antibodies isolated by the method have been shown to be functional in different immunoassays, including ELISA, immunofluorescence and Western blot. In addition, we demonstrate that by using a modified method including a negative selection step, we can isolate specific antibodies targeting the desired epitope and eliminate antibodies directed to undesired off-targets. HybriFree can be used for the reliable development of monoclonal antibodies and their subsequent production in mammalian cells. This simple protocol requires neither the culturing of B-cells nor single-cell manipulations, and only standard molecular biology laboratory equipment is needed. In principle, the method is applicable to any species for which antibody cDNA sequence information is available.

Twitter Demographics

The data shown below were collected from the profiles of 3 tweeters who shared this research output. Click here to find out more about how the information was compiled.

Mendeley readers

The data shown below were compiled from readership statistics for 71 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Japan 1 1%
Italy 1 1%
Unknown 69 97%

Demographic breakdown

Readers by professional status Count As %
Researcher 32 45%
Student > Ph. D. Student 8 11%
Student > Bachelor 8 11%
Student > Master 8 11%
Other 3 4%
Other 6 8%
Unknown 6 8%
Readers by discipline Count As %
Agricultural and Biological Sciences 25 35%
Biochemistry, Genetics and Molecular Biology 18 25%
Immunology and Microbiology 6 8%
Medicine and Dentistry 4 6%
Engineering 3 4%
Other 9 13%
Unknown 6 8%

Attention Score in Context

This research output has an Altmetric Attention Score of 5. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 03 June 2019.
All research outputs
#3,792,524
of 15,168,919 outputs
Outputs from BMC Biotechnology
#257
of 812 outputs
Outputs of similar age
#86,387
of 367,674 outputs
Outputs of similar age from BMC Biotechnology
#67
of 102 outputs
Altmetric has tracked 15,168,919 research outputs across all sources so far. This one has received more attention than most of these and is in the 74th percentile.
So far Altmetric has tracked 812 research outputs from this source. They typically receive a little more attention than average, with a mean Attention Score of 6.4. This one has gotten more attention than average, scoring higher than 67% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 367,674 tracked outputs that were published within six weeks on either side of this one in any source. This one has done well, scoring higher than 76% of its contemporaries.
We're also able to compare this research output to 102 others from the same source and published within six weeks on either side of this one. This one is in the 34th percentile – i.e., 34% of its contemporaries scored the same or lower than it.