col-79 Antibody

FAQs for Anti-CD79b Antibody Research
Below are structured FAQs addressing key scientific and methodological challenges in anti-CD79b antibody research, categorized into basic and advanced tiers.

How to validate CD79b as a therapeutic target in B-cell malignancies?

Methodological Approach:

• Expression Profiling: Use flow cytometry to quantify CD79b surface density on NHL cell lines (e.g., BJAB, Granta) and compare to normal B cells. Low expression on tumor cells (≤10% of normal B cells) does not preclude efficacy .

• Functional Assays:

  • Assess ADC internalization via confocal microscopy using pH-sensitive dyes (e.g., LysoTracker) to confirm lysosomal trafficking .

  • Measure BCR downregulation in xenograft tumors via flow cytometry (e.g., surface IgM/CD79a reduction post-ADC treatment) .

Key Data:

What in vitro models best predict anti-CD79b ADC efficacy?

Critical Factors:

• Use NHL cell lines with varying CD79b expression (e.g., DoHH2: t(14;18) translocation, Bcl2 overexpression) .

• Compare cytotoxicity of MC-MMAF (microtubule disruptor) vs. MCC-DM1 (DNA alkylator) conjugates using ATP-based viability assays .

Contradiction Note:
In vitro sensitivity does not always correlate with in vivo efficacy (e.g., DoHH2 shows modest in vitro response but strong in vivo tumor regression) .

What in vivo models are optimal for preclinical testing?

Model Selection:

• Subcutaneous xenografts (e.g., BJAB, Granta) for pharmacokinetic studies .

• Disseminated lymphoma models to mimic metastatic disease .

• hCD79 knockin mice for autoimmune applications (e.g., pristane-induced lupus, EAE) .

Dosing Protocol:

• Single 100 μg/kg dose achieves tumor regression; two doses eliminate disseminated tumors in 87.5% of mice .

How to resolve discrepancies between CD79b surface density and ADC efficacy?

Hypothesis Testing:

• Mechanistic Studies: Use immunofluorescence to track ADC trafficking to MIIC compartments, even in low-CD79b cells .

• Transcriptomic Analysis: Compare lysosomal protease (e.g., cathepsin B) activity across cell lines to identify degradation efficiency predictors .

Case Example:
Granta cells (low CD79b) show robust ADC efficacy due to efficient MIIC targeting, unlike high-CD79b lines with poor lysosomal protease activity .

What strategies enhance anti-CD79b ADC synergy with existing therapies?

Combination Approaches:

• Rituximab (anti-CD20): Co-administer with anti-CD79b ADCs to target complementary B-cell surface markers .

• BCL2 Inhibitors: Test venetoclax in follicular lymphoma models (e.g., DoHH2) to overcome apoptosis resistance .

Clinical Trial Data:

How to address ADC resistance in CD79b-low malignancies?

Experimental Design:

• CRISPR Screening: Identify resistance genes (e.g., lysosomal transporters, drug efflux pumps) in ADC-refractory clones.

• Linker Optimization: Test non-cleavable (e.g., MC-MMAF) vs. cleavable (e.g., valine-citrulline) linkers in low-pH environments .

Key Finding:
Stable-linker ADCs (MC-MMAF) outperform acid-labile conjugates (e.g., CMC-544) in CD79b-low models .

Data Contradiction Analysis