ArticleProceedings of the National Academy of Sciences of the United States of America1992
Sequencing heavy- and light-chain variable genes of single B-hybridoma cells by total enzymatic amplification.
Article in Proceedings of the National Academy of Sciences of the United States of America, 1992. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 7 papers.
What it found
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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.
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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.
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Who cites it
7 citing papers in PubMed.
- Silent development of memory progenitor B cells.Journal of immunology (Baltimore, Md. : 1950) · 2007Article
- Amplification of IgG VH and VL (Fab) from single human plasma cells and B cells.Nucleic acids research · 2000Article
- A novel strategy for generating monoclonal antibodies from single, isolated lymphocytes producing antibodies of defined specificities.Proceedings of the National Academy of Sciences of the United States of America · 1996Article
- Tracing the development of single memory-lineage B cells in a highly defined immune response.The Journal of experimental medicine · 1996Article
- Altering the antibody repertoire via transgene homologous recombination: evidence for global and clone-autonomous regulation of antigen-driven B cell differentiation.The Journal of experimental medicine · 1995Article
- Immunoglobulin A in Graves' orbital tissue: deoxyribonucleic acid amplification by polymerase chain reaction.Journal of endocrinological investigation · 1994Article
- Comparison of indirect and direct in-situ polymerase chain reaction in cell preparations and tissue sections. Detection of viral DNA, gene rearrangements and chromosomal translocations.Histochemistry · 1993Article
Corrections and comments
- Erratum issued
Authors and funding
3 authors.
Funding
Abstract
We have devised a protocol to obtain accurate and complete sequences of the immunoglobulin heavy- and light-chain variable-region (VH and VL) genes of single B-hybridoma cells that express defined V genes. The amplification achieved ranges from 2 x 10(13)- to 1 x 10(14)-fold. Only one potential Taq DNA polymerase error was observed in 7590 nucleotides of sequence, thus permitting the identification of naturally occurring somatic mutations. The two-step nature of the amplification protocol provides sufficient DNA for a minimum of 160 sets of sequencing reactions of both the VH and VL genes from one cell without cloning. The amplification of relatively long segments of DNA in the first step of the protocol permits second-step amplification and sequencing of regions that flank VH and VL codons. Fractionating cellular lysates prior to the first step of amplification permits the separate amplification of V genes on opposite sister chromatids and possibly on opposite strands of the same DNA duplex. Accurate sequencing of VH and VL genes of defined germ-line origin that are expressed by single B cells taken directly from the animal is thus made feasible by this approach.
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