About: Construction and Characterization of Single-Chain Variable Fragment Antibody Library Derived from Germline Rearranged Immunoglobulin Variable Genes

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About: Construction and Characterization of Single-Chain Variable Fragment Antibody Library Derived from Germline Rearranged Immunoglobulin Variable Genes Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : wasabi.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Construction and Characterization of Single-Chain Variable Fragment Antibody Library Derived from Germline Rearranged Immunoglobulin Variable Genes
Creator	Zhao, Q Cheng, M Au, Swn Chan, Eym Chan, Syw
Source	Medline; PMC
abstract	Antibody repertoires for library construction are conventionally harvested from mRNAs of immune cells. To examine whether germline rearranged immunoglobulin (Ig) variable region genes could be used as source of antibody repertoire, an immunized phage-displayed scFv library was prepared using splenocytic genomic DNA as template. In addition, a novel frame-shifting PCR (fsPCR) step was introduced to rescue stop codon and to enhance diversity of the complementarity-determining region 3 (CDR3). The germline scFv library was initially characterized against the hapten antigen phenyloxazolone (phOx). Sequence analysis of the phOx-selective scFvs indicated that the CDRs consisted of novel as well as conserved motifs. In order to illustrate that the diversity of CDR3 was increased by the fsPCR step, a second scFv library was constructed using a single scFv clone L3G7C as a template. Despite showing similar binding characteristics towards phOx, the scFv clones that were obtained from the L3G7C-derived antibody library gave a lower non-specific binding than that of the parental L3G7C clone. To determine whether germline library represented the endogenous immune status, specific scFv clones for nucleocapsid (N) protein of SARS-associated coronavirus (SCoV) were obtained both from naïve and immunized germline scFv libraries. Both libraries yielded specific anti-N scFvs that exhibited similar binding characteristics towards recombinant N protein, except the immunized library gave a larger number of specific anti-N scFv, and clones with identical nucleotide sequences were found. In conclusion, highly diversified antibody library can be efficiently constructed using germline rearranged immunoglobulin variable genes as source of antibody repertoires and fsPCR to diversify the CDR3.
has issue date	2011-11-11(xsd:dateTime)
bibo:doi	10.1371/journal.pone.0027406
bibo:pmid	22096568
has license	cc-by
sha1sum (hex)	23e37316e27cec6fe0e71647552b1f7b0e9255b2
schema:url	https://doi.org/10.1371/journal.pone.0027406
resource representing a document's title	Construction and Characterization of Single-Chain Variable Fragment Antibody Library Derived from Germline Rearranged Immunoglobulin Variable Genes
has PubMed Central identifier	PMC3214059
has PubMed identifier	22096568
schema:publication	PLoS One
resource representing a document's body	covid:23e37316e27cec6fe0e71647552b1f7b0e9255b2#body_text
is schema:about of	covid:arg/23e37316e27cec6fe0e71647552b1f7b0e9255b2 named entity 'scFv' named entity 'antigen' named entity 'germline' named entity 'nucleocapsid' named entity 'Antibody' named entity 'antibody' named entity 'genomic DNA' named entity 'phOx' named entity 'scFv' named entity 'protein' named entity 'germline' named entity 'Sequence analysis' named entity 'CDR3' named entity 'immunoglobulin' named entity 'hapten' named entity 'scFv' named entity 'clones' named entity 'recombinant' named entity 'Germline' named entity 'CDR3' named entity 'nucleotide sequences' named entity 'germline' named entity 'phages' named entity 'antigens' named entity 'PCR amplification' named entity 'antibody' named entity 'stop codons' named entity 'phOx' named entity 'variable region' named entity 'His-tagged' named entity 'Agarose gel' named entity 'germline' named entity 'antibody engineering' named entity 'CDR3' named entity 'CDR3' named entity 'GE Healthcare' named entity 'light chain' named entity 'denatured' named entity 'PCR' named entity 'Phage' named entity 'Perkin-Elmer' named entity 'Recombinant' named entity 'variable region' named entity 'scFv' named entity 'cloning' named entity 'CDR3' named entity 'CDR3' named entity 'immunogenic' named entity 'interleukin 11' named entity 'coronavirus' named entity 'non-specific' named entity 'CDR3' named entity 'nonspecific binding' named entity 'phOx' named entity 'scFv' named entity 'Taq polymerase' named entity 'CDR3' named entity 'Nucleotide' named entity 'antibody' named entity 'DNA fragments' named entity 'melanoma' named entity 'Applied Biosystems' named entity 'phage' named entity 'rat' named entity 'phage display library' named entity 'phagemid' named entity 'antibody' named entity 'humoral response' named entity 'antibody' named entity 'PCR' named entity 'degenerate primers' named entity 'GC-rich' named entity 'antibody' named entity 'gene' named entity 'bone marrow' named entity 'genomic DNA'

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