scholarly journals Antibody light chain variable domains and their biophysically improved versions for human immunotherapy

mAbs ◽  
2013 ◽  
Vol 6 (1) ◽  
pp. 219-235 ◽  
Author(s):  
Dae Young Kim ◽  
Rebecca To ◽  
Hiba Kandalaft ◽  
Wen Ding ◽  
Henk van Faassen ◽  
...  
Keyword(s):  
Light Chain ◽  
Variable Domains

scholarly journals The association of heavy and light chain variable domains in antibodies: implications for antigen specificity

FEBS Journal ◽  
2011 ◽  
Vol 278 (16) ◽  
pp. 2858-2866 ◽  
Author(s):  
Anna Chailyan ◽  
Paolo Marcatili ◽  
Anna Tramontano
Keyword(s):  
Light Chain ◽  
Antigen Specificity ◽  
Variable Domains

scholarly journals Recombinant immunoglobulin variable domains generated from synthetic genes provide a system for in vitro characterization of light-chain amyloid proteins

2008 ◽  
Vol 4 (3) ◽  
pp. 421-432 ◽  
Author(s):  
Priscilla Wilkins Stevens ◽  
Rosemarie Raffen ◽  
Deborah K. Hanson ◽  
Ya-Li Deng ◽  
Maria Berrios-Hammond ◽  
...  
Keyword(s):  
Light Chain ◽  
Amyloid Proteins ◽  
Synthetic Genes ◽  
In Vitro Characterization ◽  
Variable Domains

scholarly journals Design Principles for Bispecific IgGs, Opportunities and Pitfalls of Artificial Disulfide Bonds

Antibodies ◽  
2018 ◽  
Vol 7 (3) ◽  
pp. 27 ◽  
Author(s):  
Lilach Vaks ◽  
Dana Litvak-Greenfeld ◽  
Stav Dror ◽  
Galia Matatov ◽  
Limor Nahary ◽  
...  
Keyword(s):  
Disulfide Bond ◽  
Heavy Chain ◽  
Light Chain ◽  
Binding Sites ◽  
Disulfide Bonds ◽  
Optimal Solution ◽  
Bispecific Antibodies ◽  
Variable Domains ◽  
Correct Pairing ◽  
Recombinant Fragments

Bispecific antibodies (bsAbs) are antibodies with two binding sites directed at different antigens, enabling therapeutic strategies not achievable with conventional monoclonal antibodies (mAbs). Since bispecific antibodies are regarded as promising therapeutic agents, many different bispecific design modalities have been evaluated, but as many of them are small recombinant fragments, their utility could be limited. For some therapeutic applications, full-size IgGs may be the optimal format. Two challenges should be met to make bispecific IgGs; one is that each heavy chain will only pair with the heavy chain of the second specificity and that homodimerization be prevented. The second is that each heavy chain will only pair with the light chain of its own specificity and not with the light chain of the second specificity. The first solution to the first criterion (knobs into holes, KIH) was presented in 1996 by Paul Carter’s group from Genentech. Additional solutions were presented later on. However, until recently, out of >120 published bsAb formats, only a handful of solutions for the second criterion that make it possible to produce a bispecific IgG by a single expressing cell were suggested. We present a solution for the second challenge—correct pairing of heavy and light chains of bispecific IgGs; an engineered (artificial) disulfide bond between the antibodies’ variable domains that asymmetrically replaces the natural disulfide bond between CH1 and CL. We name antibodies produced according to this design “BIClonals”. Bispecific IgGs where the artificial disulfide bond is placed in the CH1-CL interface are also presented. Briefly, we found that an artificial disulfide bond between VH position 44 to VL position 100 provides for effective and correct H–L chain pairing while also preventing the formation of wrong H–L chain pairs. When the artificial disulfide bond links the CH1 with the CL domain, effective H–L chain pairing also occurs, but in some cases, wrong H–L pairing is not totally prevented. We conclude that H–L chain pairing seems to be driven by VH–VL interfacial interactions that differ between different antibodies, hence, there is no single optimal solution for effective and precise assembly of bispecific IgGs, making it necessary to carefully evaluate the optimal solution for each new antibody.

scholarly journals Formation of Amyloid Fibers by Monomeric Light Chain Variable Domains

2014 ◽  
Vol 289 (40) ◽  
pp. 27513-27525 ◽  
Author(s):  
Boris Brumshtein ◽  
Shannon R. Esswein ◽  
Meytal Landau ◽  
Christopher M. Ryan ◽  
Julian P. Whitelegge ◽  
...  
Keyword(s):  
Light Chain ◽  
Amyloid Fibers ◽  
Variable Domains

Al amyloidosis with cardiac involvement: cardiotoxicity of aggregation prone peptides deriving from variable domains of immunoglobulin light chains

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
P.E Nikolaou ◽  
G.I Nasi ◽  
I Sulaiman ◽  
P Spatharas ◽  
S Kikionis ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Cell Death ◽  
Light Chain ◽  
Hot Spots ◽  
Cytotoxic Effect ◽  
Light Chains ◽  
Al Amyloidosis ◽  
H9c2 Cells ◽  
Variable Domains

Abstract Background/Introduction Light chain (AL) amyloidosis is an uncommon malignancy manifested by systemic extracellular deposition of immunoglobulin light chain fibrils. The cardiac phenotype is characterised by ventricular wall thickening and stands as the most prominent cause of morbidity and mortality. Although, it has been established that the circulating light chains directly impair cardiomyocyte function, the cytotoxic effect of specific amyloidogenic peptides that may appear due to excessive cleavage of light chains remains unspecified. Purpose In the present work, we aimed to detect amyloidogenic “hot-spots” on the variable domains of light chains associated with cardiac AL amyloidosis (IGLV1-44 and IGLV3-01) or inferior outcomes (IGLV6-57) and define their cytotoxic effect in vitro. Methods At first, we used the curated database ALBase and we performed a multiple sequence alignment of the IGLV1-44, IGLV3-01 and IGLV6-57 inputs that derived only from patients with AL amyloidosis. “Aggregation-prone” hot-spots in the conserved amino acid sequences were identified with the aid of AMYLPRED2, a tool which combines 11 independent computational methods and provides a consensus result of potent amyloidogenic regions. Five peptides were rationally selected and synthetically produced in order to be tested in vitro. The amyloidogenic properties of the peptides were evaluated with Transmission Electron Microscopy and Congo red staining, while the rate of fibril formation at lower concentrations was monitored with Thioflavin T and confirmed with Scanning Electron Microscopy. In order to assess the cytotoxic effect of the non-polymerized peptides, H9C2 cells were incubated with the peptides for 24 hours at 200μg/mL and 100μg/mL and cell death was determined by lactate dehydrogenase release assay. Results Interestingly, sequence alignment on the variable domains of cardiac related light chains revealed the presence of several conserved domains in patients with AL amyloidosis. The chosen peptides were proven to be amyloidogenic suggesting that the variable domains share common amyloidogenic cores. Treatment of H9C2 cells with the peptides at 200μg/mL led to significant reduction in cell viability compared to vehicle treated cells (p<0.001). Two of the peptides deriving from the IGLV6-57 and IGLV3-01 significantly increased cell death at 100μg/mL (p<0.01 and p<0.001 respectively). During the 24h treatment the tested peptides comprised of soluble species and not amyloid fibrils suggesting that monomeric and oligomeric intermediates are highly toxic. Conclusion We discovered five novel amyloidogenic prone regions of cardiac related variable domains that are associated with cellular toxicity and could be exploited for targeted therapeutic interventions. Funding Acknowledgement Type of funding source: None

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