Evidence map›Paper›PMID 39447635›Full record

ArticleJournal of immunological methods2024

Human monoclonal antibody cloning and expression with overlap extension PCR and short DNA fragments.

Zachary Ende, Margarita Mishina, Robert C Kauffman, Amrita Kumar, Rashmi Kumari, Paul R Knight, Suryaprakash Sambhara

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Journal of immunological methods, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

7 authors.

Zachary EndeInfluenza Division, Centers for Disease Control and Prevention, Atlanta, GA, USA; Oak Ridge Institute for Science and Education (ORISE), CDC Fellowship Program, Oak Ridge, TN, USA.
Margarita MishinaInfluenza Division, Centers for Disease Control and Prevention, Atlanta, GA, USA.
Robert C KauffmanDepartment of Pediatrics, School of Medicine, Emory University, Atlanta, GA, USA.
Amrita KumarInfluenza Division, Centers for Disease Control and Prevention, Atlanta, GA, USA.
Rashmi KumariDepartment of Anesthesiology, Jacobs School of Medicine and Biomedical Sciences, State University of New York at Buffalo, Buffalo, NY, USA.
Paul R KnightDepartment of Anesthesiology, Jacobs School of Medicine and Biomedical Sciences, State University of New York at Buffalo, Buffalo, NY, USA.
Suryaprakash SambharaInfluenza Division, Centers for Disease Control and Prevention, Atlanta, GA, USA. Electronic address: ssambhara@cdc.gov.

Funding

Immunotherapy for acute lung injury secondary to influenzaR01HL151498 · NHLBI · STATE UNIVERSITY OF NEW YORK AT BUFFALO · PI KNIGHT III, PAUL R · 2020 to 2023
$1.7M
NHLBI NIH HHS R01 HL151498
6 · The paper itself

Abstract

Monoclonal antibodies are powerful therapeutic, diagnostic, and research tools. Methods utilized to generate monoclonal antibodies are evolving rapidly. We created a transfectable linear antibody expression cassette from a 2-h high-fidelity overlapping PCR reaction from synthesized DNA fragments. We coupled heavy and light chains into a single linear sequence with a promoter, self-cleaving peptide, and poly(A) signal to increase the flexibility of swapping variable regions from any sequence available in silico. Transfection of the linear cassette tended to generate similar levels to the two-plasmid system and generated an average of 47 μg (14-98 μg) after 5 days in 2 ml cultures with 15 unique antibody sequences. The levels of antibodies produced were sufficient for most downstream applications in less than a week. The method presented here reduces the time, cost, and complexity of cloning steps.

Indexed as

Antibodies, MonoclonalCloning, MolecularPolymerase Chain ReactionAnimalsDNAHumansImmunoglobulin Heavy ChainsImmunoglobulin Light ChainsPlasmidsTransfectionAntibodies, MonoclonalDNAImmunoglobulin Heavy ChainsImmunoglobulin Light ChainsB cellsBiolayer interferometryConfocal microscopyInfluenzaMonoclonal antibodies

Identifiers

PMID39447635
PMCPMC11585411

What OpenQuestion holds

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Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.