Sequences of the variable regions of three monoclonal antibodies specific for histidine-containing protein of the bacterial phosphoenolpyruvate:sugar phosphotransferase system

1991 ◽  
Vol 69 (4) ◽  
pp. 297-302 ◽  
Author(s):  
Teresa Steeves ◽  
M. Michele Barry ◽  
Harry W. Duckworth ◽  
E. Bruce Waygood ◽  
Jeremy S. Lee
Keyword(s):  
Monoclonal Antibodies ◽  
Gene Sequencing ◽  
Gene Families ◽  
Gene Sequences ◽  
Phosphotransferase System ◽  
Variable Regions ◽  
Gene Usage ◽  
Vh Gene

The variable regions of three monoclonal antibodies, Jel 42, Jel 44, and Jel 324, specific for the histidine-containing protein of the bacterial phosphoenolpyruvate:sugar phosphotransferase system have been sequenced from their respective mRNAs. The Vh gene families were deduced from the percent homology to the concensus gene sequences and the J gene and D gene usage was also analysed.Key words: monoclonal antibodies, gene sequencing.

Two new Ig VH gene families in Oncorhynchus mykiss

2006 ◽  
Vol 58 (11) ◽  
pp. 933-936 ◽  
Author(s):  
Gwynne D. Brown ◽  
Ilsa M. Kaattari ◽  
Stephen L. Kaattari
Keyword(s):  
Oncorhynchus Mykiss ◽  
Gene Families ◽  
Vh Gene

scholarly journals Genetic analysis of self-associating immunoglobulin G rheumatoid factors from two rheumatoid synovia implicates an antigen-driven response.

1992 ◽  
Vol 175 (3) ◽  
pp. 831-842 ◽  
Author(s):  
T Olee ◽  
E W Lu ◽  
D F Huang ◽  
R W Soto-Gil ◽  
M Deftos ◽  
...  
Keyword(s):  
Amino Acid ◽  
Heavy Chain ◽  
Immunoglobulin M ◽  
Natural Antibody ◽  
Antibody Repertoire ◽  
Rheumatoid Factors ◽  
Variable Regions ◽  
Genetic Origins ◽  
V Genes ◽  
Vh Gene

Although much has been learned about the molecular basis of immunoglobulin M (IgM) rheumatoid factors (RFs) in healthy individuals and in patients with mixed cryoglobulinemia and rheumatoid arthritis, little is known about the genetic origins of the potentially pathogenic IgG RFs in the inflamed rheumatoid synovia of patients. Recently, we generated from unmanipulated synovium B cells several hybridomas that secreted self-associating IgG RFs. To delineate the genetic origins of such potentially pathogenic RFs, we adapted the anchored polymerase chain reaction to rapidly clone and characterize the expressed Ig V genes for the L1 and the D1 IgG RFs. Then, we identified the germline counterparts of the expressed L1 IgG RF V genes. The results showed that the L1 heavy chain was encoded by a Vh gene that is expressed preferentially during early ontogenic development, and that is probably located within 240 kb upstream of the Jh locus. The overlap between this RF Vh gene and the restricted fetal antibody repertoire is reminiscent of the natural antibody-associated Vh genes, and suggests that at least part of the "potential pathogenic" IgG RFs in rheumatoid synovium may derive from the "physiological" natural antibody repertoire in a normal immune system. Indeed, the corresponding germline Vh gene for L1 encodes the heavy chain of an IgM RF found in a 19-wk-old fetal spleen. Furthermore, the comparisons of the expressed RF V genes and their germline counterparts reveal that the L1 heavy and light chain variable regions had, respectively, 16 and 7 somatic mutations, which resulted in eight and four amino acid changes. Strikingly, all eight mutations in the complementarity determining regions of the V gene-encoded regions were replacement changes, while only 6 of 11 mutations in the framework regions caused amino acid changes. Combined with L1's high binding affinity toward the Fc fragment, these results suggest strongly that the L1 IgG RF must have been driven by the Fc antigen.

scholarly journals Investigation of major amino acid residues of anti-norfloxacin monoclonal antibodies responsible for binding with fluoroquinolones

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Patamalai Boonserm ◽  
Songchan Puthong ◽  
Thanaporn Wichai ◽  
Sajee Noitang ◽  
Pongsak Khunrae ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Amino Acid ◽  
3D Structure ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Amino Acid Residues ◽  
Variable Regions ◽  
Complementarity Determining Regions ◽  
Crucial Part ◽  
Major Amino Acid

AbstractIt is important to understand the amino acid residues that govern the properties of the binding between antibodies and ligands. We studied the binding of two anti-norfloxacins, anti-nor 132 and anti-nor 155, and the fluoroquinolones norfloxacin, enrofloxacin, ciprofloxacin, and ofloxacin. Binding cross-reactivities tested by an indirect competitive enzyme-linked immunosorbent assay indicated that anti-nor 132 (22–100%) had a broader range of cross-reactivity than anti-nor 155 (62–100%). These cross-reactivities correlated with variations in the numbers of interacting amino acid residues and their positions. Molecular docking was employed to investigate the molecular interactions between the fluoroquinolones and the monoclonal antibodies. Homology models of the heavy chain and light chain variable regions of each mAb 3D structure were docked with the fluoroquinolones targeting the crucial part of the complementarity-determining regions. The fluoroquinolone binding site of anti-nor 155 was a region of the HCDR3 and LCDR3 loops in which hydrogen bonds were formed with TYR (H:35), ASN (H:101), LYS (H:106), ASN (L:92), and ASN (L:93). These regions were further away in anti-nor 132 and could not contact the fluoroquinolones. Another binding region consisting of HIS (L:38) and ASP (H:100) was found for norfloxacin, enrofloxacin, and ciprofloxacin, whereas only ASP (H:100) was found for ofloxacin.

The anti-La response of a single MRL/Mp-lpr/lpr mouse: Specificity for DNA and VH gene usage

1994 ◽  
Vol 24 (6) ◽  
pp. 1332-1338 ◽  
Author(s):  
Debra D. Bloom ◽  
E. William St. Clair ◽  
David S. Pisetsky ◽  
Stephen H. Clarke
Keyword(s):  
Gene Usage ◽  
Vh Gene

scholarly journals Ig VH gene mutational patterns indicate different tumor cell status in human myeloma and monoclonal gammopathy of undetermined significance

Blood ◽  
1996 ◽  
Vol 87 (2) ◽  
pp. 746-755 ◽  
Author(s):  
SS Sahota ◽  
R Leo ◽  
TJ Hamblin ◽  
FK Stevenson
Keyword(s):  
Multiple Myeloma ◽  
Tumor Cell ◽  
Plasma Cell ◽  
Monoclonal Gammopathy ◽  
Somatic Hypermutation ◽  
Wide Spectrum ◽  
Malignant Potential ◽  
Gene Sequences ◽  
Vh Gene ◽  
Undetermined Significance

Plasma cell tumors display a wide spectrum of clinical progression, ranging from aggressive multiple myeloma to a benign form known as monoclonal gammopathy of undetermined significance (MGUS), which requires no treatment. Because both diseases involve mature Ig- secreting plasma cells, the reason for this variation in malignant behavior is unclear. However, assessment of malignant potential is desirable for choice of treatment protocols. Ig variable (VH) gene sequences analysis has previously shown the tumor cell of multiple myeloma to be postfollicular, with mutated homogeneous clonal sequences indicating no continuing exposure to the somatic hypermutation mechanism, and this was confirmed in 7 of 7 patients. Comparison of the VH gene sequences in the monoclonal cells in MGUS yielded a different result, with 3 of 7 patients demonstrating mutated heterogeneous sequences consistent with the tumor cells remaining under the influence of the mutator. In 1 of 3 of these patients, an IgM-positive precursor cell was identified that expressed heterogeneous VH sequences similar to those of the isotype-switched plasma cell. These results indicate that the clonal cells in MGUS differ from those in myeloma and suggest that the difference may reflect malignant potential.

scholarly journals Development of Opsonic Mouse Monoclonal Antibodies against Multidrug-Resistant Enterococci

2019 ◽  
Vol 87 (9) ◽  
Author(s):  
Ermioni Kalfopoulou ◽  
Diana Laverde ◽  
Karmela Miklic ◽  
Felipe Romero-Saavedra ◽  
Suzana Malic ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Capsular Polysaccharide ◽  
Selection Process ◽  
Multidrug Resistant ◽  
Expression Vectors ◽  
Hybridoma Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bacterial Strains ◽  
Variable Regions ◽  
Content Type

ABSTRACTMultidrug-resistant enterococci are major causes of hospital-acquired infections. Immunotherapy with monoclonal antibodies (MAbs) targeting bacterial antigens would be a valuable treatment option in this setting. Here, we describe the development of two MAbs through hybridoma technology that target antigens from the most clinically relevant enterococcal species. Diheteroglycan (DHG), a well-characterized capsular polysaccharide ofEnterococcus faecalis, and the secreted antigen A (SagA), an immunogenic protein fromEnterococcus faecium, are both immunogens that have been proven to raise opsonic and cross-reactive antibodies against enterococcal strains. For this purpose, a conjugated form of the native DHG with SagA was used to raise the antibodies in mice, while enzyme-linked immunosorbent assay and opsonophagocytic assay were combined in the selection process of hybridoma cells producing immunoreactive and opsonic antibodies targeting the selected antigens. From this process, two highly specific IgG1(κ) MAbs were obtained, one against the polysaccharide (DHG.01) and one against the protein (SagA.01). Both MAbs exhibited good opsonic killing against the target bacterial strains: DHG.01 showed 90% killing againstE. faecalistype 2, and SagA.01 showed 40% killing againstE. faecium11231/6. In addition, both MAbs showed cross-reactivity toward otherE. faecalisandE. faeciumstrains. The sequences from the variable regions of the heavy and light chains were reconstructed in expression vectors, and the activity of the MAbs upon expression in eukaryotic cells was confirmed with the same immunological assays. In summary, we identified two opsonic MAbs against enterococci which could be used for therapeutic or prophylactic approaches against enterococcal infections.

Engineering Fully Human Monoclonal Antibodies from Murine Variable Regions

2010 ◽  
Vol 396 (5) ◽  
pp. 1474-1490 ◽  
Author(s):  
Matthew J. Bernett ◽  
Sher Karki ◽  
Gregory L. Moore ◽  
Irene W.L. Leung ◽  
Hsing Chen ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Human Monoclonal Antibodies ◽  
Variable Regions
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