LECRK21 Antibody
Description
Antibody Structure and Function
Antibodies are Y-shaped glycoproteins composed of two heavy and two light chains, forming variable (antigen-binding) and constant (effector function) regions . The Fab region binds antigens via complementarity-determining regions (CDRs), while the Fc region mediates immune responses . This structural framework underpins all antibody applications, including potential uses for LECRK21.
Monoclonal Antibody Engineering
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Humanization: Mouse-derived antibodies are engineered with human framework regions to reduce immunogenicity while retaining specificity .
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Affinity Optimization: Lower-affinity variants (e.g., FOLR1(Lo) TCB) may reduce off-target effects while maintaining efficacy .
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Bispecific Formats: Combining antigen-binding and immune-activating domains (e.g., T-cell bispecific antibodies) enhances therapeutic targeting .
Case Study: LECRK12 Antibody (Arabidopsis thaliana)
While LECRK21 is not explicitly documented, the following table summarizes data for LECRK12 Antibody, a related plant lectin receptor kinase antibody :
| Parameter | Details |
|---|---|
| Product Code | CSB-PA762515XA01DOA |
| Target Species | Arabidopsis thaliana (Mouse-ear cress) |
| Uniprot ID | Q7FK82 |
| Applications | Western blot, ELISA, Immunoprecipitation (assumed based on standard uses) |
| Available Sizes | 2 mL / 0.1 mL |
Research Implications:
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Lectin receptor kinases like LECRK12 regulate plant immune responses, suggesting analogous roles for LECRK21 in hypothetical studies.
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Cross-reactivity with other species or isoforms remains uncharacterized.
Validation and Quality Control
Antibody specificity is critical. Rigorous validation includes:
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KO Controls: Confirming loss of signal in knockout models (e.g., LRRK2 antibody validation) .
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Epitope Mapping: Identifying binding regions via structural assays (e.g., cryo-EM, X-ray crystallography) .
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Functional Assays: Testing neutralization efficacy (e.g., IL-21 antibody blocking cytokine activity) .
Autoimmune and Inflammatory Diseases
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Anti-CCL21 antibodies target endothelial chemokines in inflammatory bowel disease .
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Anti-TRIM21 antibodies correlate with pulmonary hypertension in systemic sclerosis .
Infectious Diseases
Challenges and Future Directions
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Specificity: Polyclonal antibodies often exhibit cross-reactivity, necessitating monoclonal alternatives .
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Glycosylation: Fc-region glycosylation impacts antibody stability and effector functions .
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Biomarker Discovery: Antibody profiling identifies clinical correlates (e.g., M1 IgM predicting severe COVID-19) .
Product Specs
Target Background
KEGG: ath:AT5G59260
STRING: 3702.AT5G59260.1
Q&A
FAQs for LECRK21 Antibody Research
Curated for academic investigators with methodological guidance and empirical data
How to validate antibody specificity for LECRK21 in experimental models?
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Methodology:
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Immunoblotting/Western blot: Use recombinant LECRK21 protein (e.g., rLipL21 ) to confirm antibody-antigen interaction. Include non-target bacterial lysates (e.g., E. coli) as negative controls .
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Cross-reactivity assays: Test against related proteins (e.g., IL-2, IL-4, IL-7) to ensure no off-target binding. For example, engineered IL-21 antibodies showed no binding to other γ-common chain cytokines .
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ELISA: Quantify binding affinity (KD) using KinExA solution equilibrium assays at physiological temperatures (37°C) .
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Table 1: Example cross-reactivity data for an anti-IL-21 antibody
| Target Protein | Binding Affinity (KD) |
|---|---|
| Human IL-21 | M |
| Mouse IL-21 | M |
| Rat IL-21 | M |
What experimental models are suitable for assessing LECRK21 antibody functionality?
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In vitro:
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In vivo:
How to resolve discrepancies in neutralizing activity across species (e.g., human vs. murine IL-21)?
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Approach:
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Structural alignment: Compare amino acid sequences of LECRK21 homologs (e.g., human vs. cynomolgus IL-21 shows 96% homology ).
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Epitope mapping: Use X-ray crystallography or hydrogen-deuterium exchange mass spectrometry (HDX-MS) to identify critical binding regions. For example, Fab fragments of antibody 362.78 revealed key IL-21 interaction sites .
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Affinity maturation: Engineer CDR regions to enhance cross-species reactivity (e.g., humanized Ab327 with modified CDRH3 ).
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Table 2: Species-specific neutralization data
| Species | IC50 (pM) | Neutralization Efficiency |
|---|---|---|
| Human | 46.7 | 6-fold > positive control |
| Cynomolgus | 41.0 | Comparable to human |
What strategies optimize antibody stability for therapeutic applications?
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Methodology:
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Constant domain engineering: Use IgG4 frameworks with stabilizing mutations (e.g., S228P in Ab327 ).
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Accelerated stability testing: Expose antibodies to stress conditions (pH 3–9, 40°C for 1 month) and assess aggregation via SEC-HPLC .
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Effector function reduction: Introduce mutations in Fc regions (e.g., L235A/G237A) to minimize ADCC/CDC activity .
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How to correlate antibody diagnostic performance with clinical disease stages?
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Case study: Anti-CCL21 antibodies in IBD :
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Immunohistochemistry: Stain mucosal biopsies from Crohn’s disease/ulcerative colitis patients to quantify CCL21 expression .
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Longitudinal monitoring: Track antibody titers in early vs. chronic infection phases (e.g., leptospirosis ).
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Receiver Operating Characteristic (ROC) analysis: Calculate sensitivity/specificity thresholds (e.g., 95% specificity for anti-rLipL21 IgG ).
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