Modification of the first constant domain of heavy chain enabled effective folding of functional anti‐forskolin antigen‐binding fragment for sensitive quantitative analysis

2019 ◽  
Vol 35 (4) ◽  
Author(s):  
Gorawit Yusakul ◽  
Seiichi Sakamoto ◽  
Hiroyuki Tanaka ◽  
Satoshi Morimoto
Keyword(s):  
Quantitative Analysis ◽  
Heavy Chain ◽  
Antigen Binding ◽  
Constant Domain

Complement activation by IgM: evidence for the importance of the third constant domain of the μ heavy chain

1987 ◽  
Vol 17 (4) ◽  
pp. 549-554 ◽  
Author(s):  
Marc J. Shulman ◽  
Catherine Collins ◽  
Nancy Pennell ◽  
Nobumichi Hozumi
Keyword(s):  
Heavy Chain ◽  
Complement Activation ◽  
The Third ◽  
Constant Domain

scholarly journals RECEPTORS ON IMMUNOCOMPETENT CELLS

1971 ◽  
Vol 134 (2) ◽  
pp. 517-531 ◽  
Author(s):  
Joseph M. Davie ◽  
Alan S. Rosenthal ◽  
William E. Paul
Keyword(s):  
Immune Response ◽  
Guinea Pig ◽  
Heavy Chain ◽  
Guinea Pigs ◽  
Bovine Serum ◽  
Antigen Binding ◽  
Agarose Beads ◽  
Cellular Immune ◽  
Draining Lymph Nodes ◽  
Antibody Secreting Cells

Guinea pigs immunized with 2,4-dinitrophenyl-guinea pig albumin (DNP-GPA) possess lymphocytes which specifically bind sufficient DNP-GPA-125I to their surface to be detected by radioautography. These lymphocytes are present in the draining lymph nodes in a frequency of ∼50/1000 lymphocytes in animals immunized 2–4 wk earlier with DNP-GPA in complete Freund's adjuvant. Nonimmunized animals have ∼0.4 DNP-GPA antigen-binding cells (ABC) per 1000 lymphocytes. An increase in the frequency of DNP-GPA ABC in peripheral blood is detectable by 5 days after immunization, which is before the time that serum anti-DNP antibody is measurable. The receptors of these ABC are hapten specific in that free ϵ-DNP-L-lysine, at low concentration, inhibits the binding of DNP-GPA-125I; DNP bovine serum alumbin (DNP-BSA) is equivalent to DNP-GPA in the inhibition of binding of DNP-GPA-125I to ABC; and both DNP-GPA agarose beads and DNP-BSA agarose beads specifically adsorb DNP-GPA-125I ABC. Anti-immunoglobulin antisera, particularly anti-γ2 sera, inhibit the binding of DNP-GPA-125I to these cells implying that the receptors are immunoglobulin, primarily of the γ2 heavy chain class. DNP-GPA-125I ABC appear to represent precursors of antibody-secreting cells and have specificity characteristics which are very different from cells, of similarly immunized guinea pigs, which mediate a cellular immune response to DNP-GPA.

Differential binding of heavy chain variable domain 3 antigen binding fragments to protein a chromatography resins

2015 ◽  
Vol 1409 ◽  
pp. 60-69 ◽  
Author(s):  
Julia Bach ◽  
Nathaniel Lewis ◽  
Kathy Maggiora ◽  
Alison J. Gillespie ◽  
Lisa Connell-Crowley
Keyword(s):  
Heavy Chain ◽  
Protein A ◽  
Variable Domain ◽  
Antigen Binding ◽  
Differential Binding ◽  
Domain 3

Binding of IgG to amyloid βA4 peptide via the heavy-chain hinge region with preservation of antigen binding

1993 ◽  
Vol 48 (2) ◽  
pp. 199-203 ◽  
Author(s):  
David Huang ◽  
Margaret Martin ◽  
Danny Hu ◽  
Allen D. Roses ◽  
Dmitry Goldgaber ◽  
...  
Keyword(s):  
Heavy Chain ◽  
Hinge Region ◽  
Antigen Binding

Highly Restricted Usage of Immunoglobulin Light Chain IGKV3-20 with Stereotyped Sequence in Primary Cold-Agglutinin Disease (CAD)

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 4137-4137 ◽  
Author(s):  
Agnieszka Malecka ◽  
Gunhild Trøen ◽  
Anne Tierens ◽  
Ingunn Østlie ◽  
Ulla Randen ◽  
...  
Keyword(s):  
Bone Marrow ◽  
B Cells ◽  
Heavy Chain ◽  
Light Chain ◽  
Cold Agglutinin ◽  
Antigen Binding ◽  
Gene Rearrangements ◽  
Cold Agglutinin Disease ◽  
Immunoglobulin Light Chain ◽  
Thermal Amplitude

Abstract Primary cold agglutinin disease (CAD) is a type of hemolytic anemia mediated by anti-I autoantibodies. Patients suffer from anemia as well as circulatory problems. However, the severity of disease differs greatly between patients. We recently demonstrated that primary CAD is caused by an underlying low grade B cell lymphoproliferative disease of the bone marrow with a typical histology that is different from lymphoplasmacytic lymphoma and, accordingly, does not display the MYD88 L265P mutation (Randen et al., Haematologica, 2013). The majority of patients display circulating monoclonal antibodies encoded by the immunoglobulin heavy chain gene IGHV4-34. The disease severity does not correlate with antibody titers, but seems to be determined by the thermal amplitude, i.e., the highest temperature at which the cold agglutinin binds to the antigen. The framework region 1 of IGHV4-34 encodes for a sequence that binds to I antigen. However, this does not explain the molecular basis of disease heterogeneity. We studied 27 patients with well-characterized primary CAD and sequenced immunoglobulin heavy as well as immunoglobulin light chains to find additional consensus regions that may determine anti-I reactivity. Bone marrow aspirates, or frozen bone marrow trephine biopsies and blood from 27 patients with well-documented primary CAD were collected. Monoclonal B cells were isolated by flow sorting (FACS Aria Ilu High speed sorter, Becton Dickinson). Viable cells were detected using the forward scatter versus side scatter dot plot. Subsequently, CD45 bright events with low side scatter features representing lymphocytes, were selected. Then, CD5 positive and CD19 negative events, i.e. T cells, were gated out using a CD5 versus CD19 dot plot leaving only B cells. Finally, monoclonal B cells were selected using the immunoglobulin light chain gate, either k or l. Clonally rearranged IGH genes were detected using the Somatic Hypermutation Assay v2.0 (Invivoscribe) and were then sequenced. Immunoglobulin light chain genes (IGL) were amplified by an in-house diagnostic protocol based on Biomed-2 primers (van Dongen et al., Leukemia, 2003). All sequences were analyzed using the IMGT database (www.imgt.org). Productive IGHV4-34 gene rearrangements were identified in 22/27 patients. In 4 patients, no productive rearrangement was identified, while in one patient a productive IGHV3-23 was seen. No significant homology of complementarity determining region 3 (CDR3) regions was found between IGHV sequences. The N-glycosylation sequence within the CDR2 region, affecting antigen-binding, was mutated in 8 patients whereas no mutations were present in 7 patients and mutations in flanking residues were seen in 6 patients. The latter mutations may modulate glycosylation efficacy. Clonal rearrangement of the IGKV3-20 was detected in 16/27 patients, clonal IGKV3-15 gene rearrangements were identified in 4/27 patients whereas other IGL genes were rearranged in 4/27 patients. No clonal IGL gene rearrangement was found in 3/27 patients. Of interest, 7 of the patients with IGKV3-20 rearrangement displayed highly homologous CDR3 regions. The latter was highly associated with an un-mutated N-glycosylation sequence of the respective IGHV4-34 sequence. In conclusion, our data show that in addition to IGHV, also IGLV usage is highly restricted in CAD. Furthermore, stereotyped IGLV sequences are seen that are mutually exclusive with mutated N-glycosylation sequences in the IGHV CDR2 sequence. These data indicate that multiple regions within the immunoglobulin heavy chain as well as immunoglobulin light chain contribute to I-antigen binding. The data suggest that subtle differences in these multiple binding sequences may contribute to the differences in thermal amplitude of I antigen binding of the antibody. The highly restricted usage of IGKV3-20 provides a rationale for vaccination with IGKV3-20 proteins, known to be immunogenic and being considered for treatment in other lymphoproliferative diseases (Martorelli et al., Clin Cancer Res, 2012). Disclosures No relevant conflicts of interest to declare.

scholarly journals Antigen-specific Proteolysis by Hybrid Antibodies Containing Promiscuous Proteolytic Light Chains Paired with an Antigen-binding Heavy Chain

2009 ◽  
Vol 284 (36) ◽  
pp. 24622-24633 ◽  
Author(s):  
Gopal Sapparapu ◽  
Stephanie A. Planque ◽  
Yasuhiro Nishiyama ◽  
Steven K. Foung ◽  
Sudhir Paul
Keyword(s):  
Heavy Chain ◽  
Light Chains ◽  
Antigen Binding
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