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Localization of Therapeutic Fab-CHP Conjugates to Sites of Denatured Collagen for the Treatment of Rheumatoid Arthritis
journal contribution
posted on 2020-07-20, 18:05 authored by Keith
J. Arlotta, Boi Hoa San, Hong-Hua Mu, S. Michael Yu, Shawn C. OwenRheumatoid
arthritis (RA) is an autoimmune disease characterized
by chronic inflammation in synovial joints and protease-induced cartilage
degradation. Current biologic treatments for RA can effectively reduce
symptoms, primarily by neutralizing the proinflammatory cytokine TNFα;
however, continued, indiscriminate overinhibition of inflammatory
factors can significantly weaken the host immune system, leading to
opportunistic infections and interrupting treatment. We hypothesize
that localizing anti-TNFα therapeutics to denatured collagen
(dCol) present at arthritic joints, via conjugation with collagen-hybridizing
peptides (CHPs), will reduce off-site antigen binding and maintain
local immunosuppression. We isolated the antigen-binding fragment
of the clinically approved anti-TNFα therapeutic infliximab
(iFab) and prepared iFab-CHP conjugates via lysine-based conjugation
with an SMCC linker. After successful conjugation, confirmed by LC-MS,
the binding affinity of iFab-CHP was characterized by ELISA-like assays,
which showed comparable antigen binding relative to infliximab, comparable
dCol binding relative to CHP, and the hybrid ability to bind both
dCol and TNFα simultaneously. We further demonstrated localization
of Fab-CHP to areas of high dCol in vivo and promising therapeutic
efficacy, assessed by histological staining (Safranin-O and H&E),
in a pilot mouse study.
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Keywords
antigen-binding fragmentTNF αhistological stainingLC-MSlysine-based conjugationoff-site antigen bindingDenatured Collagenlocalizing anti-TNF α therapeuticscollagen-hybridizing peptidesSMCC linkerdCol bindingantigen bindinganti-TNF αiFab-CHP conjugatesRAELISA-like assayssynovial jointspilot mouse studybinding affinityTherapeutic Fab-CHP ConjugatesinfliximabRheumatoid Arthritis Rheumatoid art...denatured collagenprotease-induced cartilage degradation
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