RPL35A Antibody

What is RPL35A and why is it significant in scientific research?

RPL35A (60S ribosomal protein L35a) is a ribosomal protein that functions as an essential component of the large ribosomal subunit. This protein plays critical roles in 60S ribosomal subunit formation and is required for the proliferation and viability of hematopoietic cells . The significance of RPL35A extends beyond its structural role in ribosomes, as it has been found to bind to both initiator and elongator tRNAs, suggesting functional involvement at the P site or both P and A sites during translation .

RPL35A gained particular research interest when mutations in its gene were identified in Diamond-Blackfan anemia (DBA), an inherited bone marrow failure syndrome characterized by anemia, congenital abnormalities, and cancer predisposition . Furthermore, recent studies have implicated RPL35A in cancer progression, particularly in ovarian cancer where it promotes the binding of transcription factor YY1 to the CTCF promoter . These diverse biological functions make RPL35A antibodies invaluable tools for investigating ribosome biology, hematological disorders, and cancer mechanisms.

What applications are RPL35A antibodies most commonly used for in scientific research?

RPL35A antibodies are versatile research tools employed in multiple experimental contexts:

• Immunohistochemistry (IHC): Used to detect RPL35A protein expression patterns in tissue sections, enabling correlation with clinical parameters in pathological conditions .

• Immunofluorescence (IF) and Immunocytochemistry (ICC): Allow visualization of RPL35A subcellular localization and potential co-localization with interacting partners .

• Flow Cytometry (FACS): Particularly useful with fluorophore-conjugated RPL35A antibodies (such as APC-conjugated versions) for quantitative analysis of RPL35A expression in different cell populations .

• Enzyme-Linked Immunosorbent Assay (ELISA/EIA): Used for quantitative measurement of RPL35A protein levels in biological samples .

• Chromatin Immunoprecipitation (ChIP): Essential for investigating the involvement of RPL35A in transcriptional regulation, as demonstrated in studies of ovarian cancer where RPL35A influences the binding of transcription factors to promoter regions .

These diverse applications highlight why RPL35A antibodies are indispensable tools in both basic research and translational studies investigating ribosome biology, hematological disorders, and cancer mechanisms.

How should researchers validate RPL35A antibodies for experimental use?

Proper validation of RPL35A antibodies is critical for ensuring experimental reliability. A comprehensive validation approach should include:

• Specificity Testing:

  • Western blot analysis showing a single band at the expected molecular weight of RPL35A (approximately 12.5 kDa)

  • Positive and negative control tissues/cell lines with known RPL35A expression levels

  • Blocking peptide competition assays to confirm specific binding

  • Knockdown/knockout validation using RPL35A-specific shRNA or CRISPR-Cas9, as demonstrated in studies where shRNA inhibition of RPL35A affected maturation of 28S and 5.8S rRNAs

• Cross-Reactivity Assessment:

  • Testing against closely related ribosomal proteins

  • Evaluation across relevant species if cross-species reactivity is claimed (human RPL35A antibodies are most common in research settings)

• Application-Specific Validation:

  • For IHC: Optimize fixation methods, antigen retrieval, and antibody concentration

  • For IF/ICC: Confirm subcellular localization patterns consistent with known RPL35A distribution

  • For ChIP applications: Validate enrichment at known binding sites using qPCR with specific primers (e.g., for the CTCF promoter as shown in ovarian cancer research)

• Batch-to-Batch Consistency:

  • Compare new antibody lots with previously validated lots

  • Maintain positive control samples for standardization

Thoroughly validated antibodies, such as affinity-purified rabbit polyclonal antibodies against human RPL35A (amino acids 1-110) expressed in E. coli, have demonstrated reliable performance across multiple experimental applications .

What is the connection between RPL35A and Diamond-Blackfan anemia (DBA)?

The connection between RPL35A and Diamond-Blackfan anemia (DBA) represents a significant breakthrough in understanding the molecular basis of this rare inherited bone marrow failure syndrome:

• Discovery Context:
  RPL35A was identified as a DBA-associated gene through a candidate gene approach combining high-resolution genomic mapping and gene expression microarray analysis of DBA patients with chromosome 3q deletions . This discovery expanded the understanding of DBA beyond previously identified small ribosomal subunit gene mutations (RPS19, RPS24, RPS17).

• Mutational Spectrum:
  Multiple types of pathogenic variants in RPL35A have been identified in DBA patients:

  • Large deletions involving the entire RPL35A gene

  • Loss-of-function (LOF) variants: splice site defects, nonsense mutations, frameshift mutations

  • Missense mutations and inframe deletions

• Genotype-Phenotype Correlations:
  A multi-institutional study of 45 DBA patients with pathogenic RPL35A variants revealed distinct clinical patterns:

  RPL35A Variant Type	Clinical Features	Treatment Response
  Large deletions	More severe phenotype, neutropenia (70%), immune system abnormalities, recurrent infections	Often steroid-resistant, transfusion-dependent
  LOF variants	Earlier age at diagnosis of anemia	Generally better response to treatment
  Missense/inframe deletions	Less severe phenotype	Better response to steroids

  These differences were statistically significant (P<0.01) when comparing large deletions to other pathogenic variants .

• Molecular Mechanism:
  Functional studies using shRNA inhibition demonstrated that RPL35A is essential for:

  • Maturation of 28S and 5.8S rRNAs

  • 60S ribosomal subunit biogenesis

  • Normal cell proliferation and survival

• Diagnostic Implications:
  Erythrocyte adenosine deaminase (eADA) values, a diagnostic marker for DBA, are elevated in both patients with large deletions and those with other RPL35A variants, with no significant difference between groups (P=0.682) .

This established connection between RPL35A and DBA underscores the importance of ribosome biogenesis in hematopoiesis and provides a mechanistic framework for investigating other ribosomopathies.

How can RPL35A antibodies be utilized to study altered ribosome biogenesis in disease models?

RPL35A antibodies offer powerful tools for investigating ribosome biogenesis abnormalities in disease models, particularly for Diamond-Blackfan anemia (DBA) and cancer research. Implementation strategies include:

• Polysome Profiling with Immunoblotting:

  • Fractionate cellular lysates on sucrose gradients to separate free ribosomal subunits, monosomes, and polysomes

  • Analyze fractions by Western blotting using RPL35A antibodies to track incorporation into pre-60S subunits, mature 60S subunits, and active ribosomes

  • Compare profiles between wild-type and disease models to identify defects in large subunit assembly

  • This approach can reveal whether RPL35A mutations affect incorporation into ribosomes or destabilize the entire 60S subunit

• Ribosome Biogenesis Intermediates Characterization:

  • Employ RPL35A antibodies in immunoprecipitation to isolate pre-ribosomal particles

  • Analyze co-precipitated rRNA species (particularly 28S and 5.8S rRNAs) and protein components

  • Use pulse-chase labeling with metabolic RNA labeling to track rRNA processing kinetics

  • Apply this methodology to compare normal cells with those carrying RPL35A mutations or depletion, as studies have demonstrated that RPL35A is essential for maturation of 28S and 5.8S rRNAs

• Nucleolar Stress Response Assessment:

  • Utilize immunofluorescence with RPL35A antibodies to monitor nucleolar morphology and protein localization

  • Track redistribution of RPL35A under ribosomal stress conditions

  • Investigate interactions with p53 pathway components, as ribosomal protein imbalance often activates p53-dependent cell cycle arrest

• Translation Fidelity Analysis:

  • Study translation dynamics in cells with altered RPL35A function using reporter systems

  • Employ RPL35A antibodies in ribosome profiling experiments to analyze ribosome positioning and translation efficiency at a genome-wide level

  • Investigate connections between RPL35A's role in binding both initiator and elongator tRNAs at the P site or P and A sites

• Integrated -Omics Approach:

  • Combine RPL35A antibody-based proteomics (immunoprecipitation-mass spectrometry) with transcriptomics, especially in DBA patient-derived cells

  • Profile alterations in ribosome composition and associated factors

  • Link ribosome biogenesis defects to downstream consequences on specific mRNA translation

These methodological approaches using RPL35A antibodies facilitate a deeper understanding of how ribosome biogenesis perturbations contribute to disease pathogenesis, potentially revealing therapeutic vulnerabilities.

What are the best experimental designs for investigating RPL35A's role in cancer progression?

Research has revealed that RPL35A drives ovarian cancer progression through specific transcriptional regulatory mechanisms . To thoroughly investigate RPL35A's role in cancer, consider these optimized experimental designs:

• Gene Expression Modulation Studies:

  • Knockdown Design: Use multiple shRNA constructs targeting different regions of RPL35A (both coding sequence and 3' UTR) with appropriate controls (e.g., luciferase shRNA)

  • Overexpression Strategy: Employ inducible expression systems to avoid selection against potential growth inhibition

  • Rescue Experiments: Reintroduce shRNA-resistant RPL35A variants to confirm phenotype specificity

  • Primary Endpoint Measurements: Cell proliferation, migration, invasion, and apoptosis assessments with standardized assays

• ChIP-seq and ChIP-qPCR Workflows:

  • Sample Preparation: Fix cells with formaldehyde, lyse in SDS buffer, and fragment DNA by sonication

  • Immunoprecipitation Controls: Include negative control (normal mouse IgG), positive control (Histone H3), and specific antibody (anti-YY1)

  • Promoter Analysis: Design specific primers for target promoters (e.g., CTCF promoter)

  • Primer Design Example: For CTCF promoter: 5′-CCCAAGTTTATCACACCGCTCA-3′ and 5′-AAGGCAGCATCTAGGAAGTCATG-3′

• Mechanistic Interaction Studies:

  • Co-immunoprecipitation: Use RPL35A antibodies to pull down protein complexes and identify interacting partners

  • Proximity Ligation Assays: Visualize and quantify protein-protein interactions in situ

  • RIME (Rapid Immunoprecipitation Mass spectrometry of Endogenous proteins): Identify chromatin-associated RPL35A interactome

  • Sequential ChIP: Determine co-occupancy of RPL35A with transcription factors like YY1 at target promoters

• Translational Research in Patient Samples:

  • Tissue Microarray Analysis: Use RPL35A antibodies for immunohistochemical evaluation of expression in tumor versus normal tissues

  • Correlation Studies: Analyze associations between RPL35A expression and clinical parameters including TNM staging and patient survival

  • Expression Quantification: Employ qRT-PCR with validated primers (e.g., RPL35A forward: 5′-GAAGGTGTTTACGCCCGAGAT-3′ and reverse: 5′-CGAGTTACTTTTCCCCAGATGAC-3′)

• In Vivo Models:

  • Xenograft Studies: Compare tumor growth, metastasis, and response to therapy between RPL35A-modulated cancer cells and controls

  • Patient-Derived Xenografts: Test RPL35A-targeting strategies in models that better recapitulate human tumor heterogeneity

  • Assessment Parameters: Tumor volume, metastatic burden, immunohistochemical analysis of proliferation and apoptosis markers

These experimental designs provide a comprehensive framework for investigating RPL35A's oncogenic functions, particularly its role in promoting transcription factor binding to target promoters in cancer progression.

How can researchers differentiate between canonical and non-canonical functions of RPL35A in their experiments?

Distinguishing between RPL35A's canonical role in ribosome structure/function and its emerging non-canonical functions presents a significant challenge in research. Here are methodological approaches to differentiate these functions:

• Subcellular Fractionation Analysis:

  • Methodology: Separate cellular compartments (cytoplasmic, nuclear, nucleolar, chromatin-bound)

  • Detection: Use RPL35A antibodies for Western blotting of each fraction

  • Interpretation: Non-ribosomal pools of RPL35A in unexpected locations (nuclear, chromatin-bound) suggest non-canonical functions

  • Controls: Include markers for each compartment (e.g., nucleolin for nucleolus, histone H3 for chromatin)

• Mutational Analysis Strategy:

  • Design: Create RPL35A variants with mutations that specifically affect:
    a) Ribosome incorporation (structural interface mutations)
    b) tRNA binding (P-site interaction mutations)
    c) Protein-protein interaction sites (surface-exposed regions)

  • Functional Testing: Assess which cellular functions are disrupted by each mutation type

  • Readouts: Ribosome assembly (polysome profiles), translation (metabolic labeling), and non-canonical functions (e.g., transcription factor binding)

• Temporal Dynamics Investigation:

  • Experimental Approach: Use rapid RPL35A depletion systems (e.g., auxin-inducible degron)

  • Time-Course Analysis: Determine which functions are affected first

  • Hypothesis: Canonical functions requiring new ribosome assembly may take longer to manifest than direct non-canonical functions

  • Measurement: Monitor ribosome biogenesis (28S and 5.8S rRNA maturation) versus transcription factor binding kinetics

• Protein Interaction Network Mapping:

  • Methodology: Combine immunoprecipitation with mass spectrometry (IP-MS)

  • Comparative Analysis: Identify interactors in different cellular compartments

  • Bioinformatic Classification: Group interactors into ribosome-related versus non-ribosomal proteins

  • Validation: Confirm key interactions with co-IP and proximity ligation assays

• Ribosome-Free RPL35A Population Studies:

  • Isolation Strategy: Immunoprecipitate RPL35A after ribosome depletion (e.g., by ultracentrifugation)

  • Functional Characterization: Assess activities of this pool specifically

  • Associated Proteins: Identify unique partners of non-ribosomal RPL35A

  • Chromatin Association: Use ChIP-seq to map genome-wide binding sites of the non-ribosomal fraction

• Comparison Table: Experimental Features for Distinguishing RPL35A Functions

By systematically applying these approaches, researchers can delineate the diverse functions of RPL35A and understand how they may independently contribute to normal physiology and disease states.

What are the optimal conditions for using RPL35A antibodies in ChIP and ChIP-seq experiments?

Chromatin immunoprecipitation (ChIP) experiments using RPL35A antibodies require careful optimization to investigate its role in transcriptional regulation, particularly in cancer contexts. Based on successful implementations in ovarian cancer research , here are the recommended protocols and considerations:

• Sample Preparation Protocol:

  • Crosslinking: Fix cells with 1% formaldehyde for 10 minutes at room temperature

  • Quenching: Add glycine to a final concentration of 0.125M

  • Cell Lysis: Use SDS lysis buffer (1% SDS, 10mM EDTA, 50mM Tris-HCl, pH 8.0)

  • Chromatin Fragmentation: Sonicate to generate DNA fragments of 200-500bp

    • Optimize sonication conditions (amplitude, pulse duration, cycle number) for each cell type

    • Verify fragment size by agarose gel electrophoresis

• Antibody Selection and Validation:

  • Recommended Antibody Type: Affinity-purified rabbit polyclonal antibodies against human RPL35A

  • Pre-clearing Step: Incubate chromatin with protein A/G beads and non-immune IgG

  • Control Antibodies: Include in parallel experiments:

    • Negative control: normal rabbit/mouse IgG

    • Positive control: anti-Histone H3

• ChIP Protocol Optimization:

  • Antibody Amount: Titrate between 2-5μg per ChIP reaction

  • Chromatin Amount: Use 25-50μg of chromatin per reaction

  • Incubation Conditions: Rotate overnight at 4°C

  • Washing Stringency: Sequential washes with increasing salt concentration

  • Elution and Reversal of Crosslinks: 65°C incubation for 4-6 hours

  • DNA Purification: Column-based methods for highest recovery

• ChIP-qPCR Analysis:

  • Primer Design: Create primers spanning predicted binding sites:

    • Example for CTCF promoter: 5′-CCCAAGTTTATCACACCGCTCA-3′ and 5′-AAGGCAGCATCTAGGAAGTCATG-3′

  • qPCR Conditions: Use SYBR-based detection with appropriate controls

  • Data Normalization: Express results as percent input or fold enrichment over IgG control

  • Biological Replicates: Minimum of three independent experiments

• ChIP-seq Considerations:

  • Library Preparation: Use low-input library preparation kits for potentially limited ChIP material

  • Sequencing Depth: Minimum 20 million uniquely mapped reads per sample

  • Peak Calling: Employ MACS2 or similar algorithms with appropriate parameters

  • Data Validation: Confirm selected peaks by ChIP-qPCR

  • Motif Analysis: Identify enriched DNA motifs within peak regions using MEME-ChIP or similar tools

• Differential Binding Analysis in Experimental Conditions:

  • Comparison Strategy: Analyze RPL35A binding in control versus experimental conditions (e.g., overexpression models)

  • Integration with Gene Expression: Correlate binding changes with differential gene expression

  • Pathway Enrichment: Analyze biological pathways associated with differential binding sites

These optimized conditions have successfully revealed RPL35A's role in promoting the binding of transcription factor YY1 to the CTCF promoter in ovarian cancer , highlighting the effectiveness of this approach for investigating non-canonical functions of RPL35A in transcriptional regulation.

How can researchers effectively use RPL35A antibodies to study its role in Diamond-Blackfan anemia (DBA) patient samples?

Studying RPL35A in Diamond-Blackfan anemia (DBA) patient samples presents unique challenges due to the rarity of the disease and the diverse spectrum of pathogenic variants. The following methodological framework maximizes the research value of RPL35A antibodies in this context:

• Patient Sample Classification and Analysis:

  • Stratify Samples by mutation type:

    • Large deletions involving entire RPL35A gene

    • Loss-of-function variants (splice site, nonsense, frameshift)

    • Missense mutations or inframe deletions

  • Sampling Considerations: Collect peripheral blood, bone marrow aspirates, and when available, skin fibroblasts (for establishment of patient-derived cell lines)

  • Control Selection: Include age-matched healthy donors and DBA patients with mutations in other ribosomal protein genes (e.g., RPS19)

• Expression Analysis Protocol:

  • Western Blot Optimization:

    • Use lymphoblastoid cell lines (LCLs) derived from patients

    • Extract proteins using RIPA buffer with protease inhibitors

    • Load equal protein amounts (20-50μg) per lane

    • Recommended antibody dilution: 1:1000 for primary RPL35A antibody

    • Quantify relative expression normalized to housekeeping proteins

  • Immunohistochemistry for bone marrow biopsies:

    • Formalin-fixed paraffin-embedded (FFPE) tissue processing

    • Antigen retrieval in citrate buffer (pH 6.0)

    • Use RPL35A antibodies optimized for IHC applications

    • Counterstain with hematoxylin for nuclear visualization

• Ribosome Biogenesis Assessment:

  • Northern Blot Protocol for pre-rRNA processing:

    • Extract total RNA from patient cells

    • Resolve on formaldehyde-agarose gels

    • Probe for pre-rRNA species (28S, 5.8S)

    • Compare processing patterns between patient groups

  • Polysome Profiling:

    • Prepare cytoplasmic extracts from patient-derived cells

    • Fractionate on 10-50% sucrose gradients

    • Analyze 40S, 60S, 80S, and polysome peaks

    • Compare profiles between different RPL35A variant types

• Functional Studies in Patient-Derived Cells:

  • Proliferation and Apoptosis Assays:

    • Culture erythroid progenitors from patient bone marrow

    • Monitor cell growth, cell cycle progression, and apoptosis

    • Compare results across different RPL35A mutation types

  • Erythroid Differentiation Analysis:

    • Culture CD34+ cells from patient samples

    • Induce erythroid differentiation

    • Monitor expression of differentiation markers using flow cytometry

    • Correlate differentiation capacity with RPL35A variant type

• Rescue Experiments:

  • Lentiviral Transduction of wild-type RPL35A into patient cells

  • Readouts: Pre-rRNA processing, polysome profiles, cell proliferation

  • Hypothesis Testing: Determine if different RPL35A variants show variable rescue efficiency

  • Correlation: Link molecular phenotypes with clinical severity (e.g., transfusion dependence, response to steroids)

• Comparative Analysis Table: RPL35A Function in Different DBA Patient Groups

These comprehensive methodological approaches enable researchers to establish genotype-phenotype correlations and understand the molecular basis of clinical variability in DBA patients with different types of RPL35A mutations .

What emerging technologies could enhance the utility of RPL35A antibodies in ribosome research?

Several cutting-edge technologies are poised to revolutionize how RPL35A antibodies can be utilized in ribosome biology research:

• Proximity-Based Labeling Combined with Proteomics:

  • BioID/TurboID Technology: Fuse RPL35A with biotin ligase to identify proteins in close proximity within living cells

  • Implementation Strategy: Generate stable cell lines expressing RPL35A-BioID fusion proteins

  • Application Advantage: Maps dynamic interaction networks in different cellular compartments

  • Research Question Example: Identify differential RPL35A proximity interactomes in normal versus disease states

• Super-Resolution Microscopy Techniques:

  • STORM/PALM Applications: Use fluorophore-conjugated RPL35A antibodies for single-molecule localization microscopy

  • Experimental Design: Co-localization studies with markers for different nuclear bodies and cytoplasmic ribosomes

  • Resolution Improvement: Visualize RPL35A distribution at 20-30nm resolution versus 200-300nm with conventional microscopy

  • Key Investigation: Track RPL35A movement between nucleolus, nucleoplasm, and cytoplasm during ribosome biogenesis

• Cryo-Electron Tomography with Gold-Labeled Antibodies:

  • Method Development: Conjugate RPL35A antibodies with gold nanoparticles

  • Application: Visualize RPL35A positioning within intact cellular ribosomes

  • Advantage: Preserves native cellular context unlike traditional cryo-EM of purified ribosomes

  • Research Goal: Map structural changes in ribosomes from cells with RPL35A mutations

• Single-Cell Protein Analysis Technologies:

  • Mass Cytometry (CyTOF): Use metal-tagged RPL35A antibodies for high-dimensional analysis

  • Single-Cell Western Blotting: Quantify RPL35A expression heterogeneity

  • Experimental Context: Analyze primary bone marrow samples from DBA patients

  • Research Question: How does RPL35A expression vary within erythroid progenitor subpopulations?

• CRISPR-Based Genomic Screens Combined with RPL35A Antibodies:

  • Methodology: Conduct genome-wide CRISPR screens for modifiers of RPL35A function

  • Readout: Use RPL35A antibodies to track changes in expression, localization, or interaction partners

  • Application: Identify novel factors influencing RPL35A's role in ribosome biogenesis or cancer progression

  • Potential Discovery: New therapeutic targets for ribosomopathies or RPL35A-driven cancers

• Spatial Transcriptomics with Protein Detection:

  • Technical Approach: Combine in situ RNA sequencing with RPL35A immunofluorescence

  • Implementation: Apply to tissue sections from normal and disease samples

  • Research Value: Correlate RPL35A protein levels with spatial transcriptome alterations

  • Example Application: Map transcript-specific translation efficiency differences in RPL35A-mutated cells

• Ribosome Profiling with RPL35A-Specific Immunoprecipitation:

  • Method Innovation: Selectively isolate RPL35A-containing ribosomes before ribosome profiling

  • Hypothesis Testing: Determine if RPL35A-containing ribosomes preferentially translate specific mRNA subsets

  • Experimental Context: Compare wild-type cells with those expressing mutant RPL35A variants found in DBA

  • Potential Finding: RPL35A mutations may cause selective translation defects rather than global impairment

These emerging technologies promise to deepen our understanding of RPL35A's multifaceted roles in ribosome biology, hematopoiesis, and cancer, potentially revealing new therapeutic targets for RPL35A-associated diseases.

How might RPL35A serve as a therapeutic target in cancer, and what research tools would be needed?

Recent research demonstrating RPL35A's role in ovarian cancer progression suggests its potential as a therapeutic target. Developing this potential requires specialized research tools and methodological approaches:

• Therapeutic Target Validation Strategy:

  • Conditional Knockdown Models: Develop inducible shRNA or doxycycline-regulated CRISPR systems targeting RPL35A

  • Xenograft Studies: Assess tumor growth inhibition following RPL35A depletion in established tumors

  • Patient-Derived Organoids: Test RPL35A targeting in 3D cultures preserving tumor heterogeneity

  • Therapeutic Window Assessment: Compare effects of partial RPL35A inhibition in cancer versus normal cells

• Druggable Interface Identification:

  • Structural Biology Approaches:

    • Generate high-resolution structures of RPL35A-YY1 and RPL35A-CTCF interactions

    • Map binding interfaces using site-directed mutagenesis

    • Design peptide mimetics to disrupt protein-protein interactions

  • Antibody-Based Mapping:

    • Develop domain-specific RPL35A antibodies to identify functional regions

    • Use epitope mapping to pinpoint interaction surfaces

    • Test antibody fragments for disruption of cancer-promoting interactions

• Small Molecule Screening Platforms:

  • High-Throughput Screening Design:

    • Develop fluorescence resonance energy transfer (FRET) assays for RPL35A-YY1 interaction

    • Establish cell-based reporter systems for CTCF promoter activity

    • Screen compound libraries against these targets

  • Validation Methodologies:

    • Confirm hit compounds using surface plasmon resonance

    • Verify mechanism of action with RPL35A antibody-based assays

    • Test effects on transcriptional regulation using ChIP-qPCR

• Therapeutic Antibody Development:

  • Approach: Generate function-blocking antibodies targeting non-ribosomal RPL35A

  • Screening Strategy: Test antibody panels for inhibition of RPL35A-YY1 binding

  • Delivery Methods: Explore antibody-drug conjugates or cell-penetrating antibody fragments

  • Efficacy Testing: Measure inhibition of cancer cell proliferation and migration

• Biomarker Development for Patient Stratification:

  • IHC Protocols: Standardize RPL35A immunohistochemistry for clinical samples

  • Expression Thresholds: Establish cutoffs for "RPL35A-high" tumors

  • Companion Diagnostics: Develop assays for RPL35A-dependent pathways

  • Patient Selection: Identify candidates most likely to benefit from RPL35A-targeted therapy

• Combination Therapy Exploration:

  • Synergy Screening: Test RPL35A inhibition with standard chemotherapeutics

  • Mechanistic Investigations: Use RPL35A antibodies to track changes in signaling pathways

  • Rational Combinations: Target both ribosomal and non-ribosomal functions of RPL35A

  • Resistance Mechanisms: Monitor adaptations to RPL35A targeting using proteomics

• Predictive Response Table: Potential Therapeutic Strategies Based on RPL35A Function

The development of these specialized research tools would establish RPL35A as a validated cancer therapeutic target and provide the necessary framework for translating biological insights into clinical applications.