LCR65 Antibody
Description
GPR65 Antibody Overview
GPR65 antibodies are immunological tools designed to detect and study the GPR65 receptor, which plays roles in immune regulation, acid-sensing, and cancer immunotherapy. These antibodies enable:
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Detection of GPR65 expression in immune cells (e.g., NK cells, monocytes) and tumor microenvironments .
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Functional studies of GPR65's role in cAMP/PKA signaling, lysosomal homeostasis, and T-cell inhibition .
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Therapeutic development for inflammatory diseases and cancer .
Antibody Development and Validation
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Chimeric antigen scaffolds displaying GPR65 extracellular loops (ECL1, ECL2) were used to generate phage-derived antibodies. Three clones (Ab1.1, Ab5.2, Ab5.3) demonstrated binding to native GPR65 on HEK cells and primary immune cells .
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Specificity: Antibodies showed preferential binding to CD16+ NK cells, B cells, and monocytes, but not T cells .
Functional Insights
Mechanistic Roles of GPR65
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Immune Regulation:
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Therapeutic Targeting:
Technical Considerations
Product Specs
Composition: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
FAQs for LCR65 Antibody in Academic Research
What advanced methodologies resolve cross-reactivity challenges with LCR65 antibodies in infectious disease studies?
In leprosy research, LCR65 antibodies targeting Mycobacterium leprae’s 65-kD antigen (rML65) face cross-reactivity with homologous bacterial proteins. Solutions include:
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Epitope mapping: Use peptide arrays to identify linear vs. conformational binding sites .
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Cellular vs. humoral assays: Pair antibody titers (ELISA) with T-cell proliferation tests + IL-2 supplementation to distinguish adaptive immune responses .
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Blocking strategies: Pre-adsorb antibodies with recombinant proteins from related species (e.g., M. tuberculosis) to isolate M. leprae-specific signals .
Key finding: Inverse correlation exists between anti-rML65 IgG levels (high in lepromatous leprosy) and T-cell reactivity (low), necessitating dual-platform validation .
How can LCR65 antibodies be integrated into multiplex assays for biomarker discovery?
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Panel design: Combine LCR65 with antibodies against glial fibrillary acidic protein (GFAP) or neurofilament light chain (NfL) for neurodegenerative biomarker panels .
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Platform optimization:
Data table:
| Target | Assay Type | Sensitivity (LOD) | Cross-Reactivity Risk |
|---|---|---|---|
| p-α-syn (PD) | Immunogold EM | 0.1 ng/mL | Non-phospho α-syn |
| rML65 (Leprosy) | IFN-γ ELISpot | 10 spot-forming units | M. tuberculosis |
What statistical approaches address variability in LCR65 antibody performance across cohorts?
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Finite mixture modeling: Segregate seropositive/seronegative populations in antibody titer datasets, accounting for outliers (e.g., median vs. mean in skewed distributions) .
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Multivariate regression: Adjust for confounders like age or HLA haplotypes in leprosy endemic populations .
Example: A study of 15 antibodies showed median titers for LCR65 varied widely (Q1-Q3: 4–32 vs. 8–64 in alternative assays), highlighting the need for non-parametric statistical tests .
How do novel AI-driven platforms enhance LCR65 antibody engineering?
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CDRH3 design: Generative AI models (e.g., IgLM) create synthetic CDRH3 sequences with germline V/J templates, improving affinity for targets like viral antigens .
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Structural validation: ImmuneBuilder predicts paratope-epitope docking for SARS-CoV-2 cross-reactive antibodies, a framework applicable to LCR65 .
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High-throughput screening: Couple AI-designed libraries with phage display to isolate high-affinity clones against rare epitopes .
Case study: AI-generated anti-SARS-CoV-2 antibodies achieved broad HA binding despite low sequence homology to natural clonotypes .
What controls are essential when using LCR65 antibodies in immunohistochemistry?
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Tissue controls: Include PD patient amygdala sections (positive) and α-syn knockout models (negative) .
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Preabsorption controls: Incubate antibodies with excess phosphorylated peptide to confirm signal loss .
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Isotype controls: Use non-specific IgG to rule out Fc-mediated binding in inflamed tissues .
Pitfall: Mitochondrial co-localization in neuronal cells may require dual staining with TOMM20 (mitochondrial marker) .
How do long-lived plasma cells (LLPCs) confound LCR65 antibody persistence studies in transplantation?
LLPCs secrete alloantibodies independent of B-cell depletion therapies. Strategies to address this:
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Proteasome inhibitors: Bortezomib reduces LLPC reservoirs in antibody-mediated kidney rejection .
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Dual-target CAR-T cells: Engineered to deplete CD19+ B cells and CD138+ plasma cells in preclinical models .
Data insight: Anti-thymocyte globulin (ATG) reduces LLPCs by 60% in sensitized patients, but rebound occurs post-treatment .
