Customize ADF9 Antibody

Description

ADK9 Antibody: Functional Characterization

ADK9 is a monoclonal antibody (MAb) targeting AAV9 capsids, developed to address preexisting immunity challenges in gene therapy. Key features include:

Property	Details	Source
Target Epitope	Binds near icosahedral 3-fold axes of AAV9 capsid, involving residues S454 and P659
Neutralization Capacity	Reduces AAV9 transduction by 100- to 1,000-fold in spleen, heart, and liver tissues
Clinical Relevance	Neutralizing antibodies like ADK9 exclude ~40–80% of patients from AAV9-based gene therapies

Seroprevalence and Clinical Impact

A retrospective study of 882 pediatric SMA patients showed:

Elevated AAV9 Antibodies: 13.04% at first test, decreasing with age (18.2% in infants ≤3 months vs. <5% after 9 months) .
Gender Disparity: Higher prevalence in males (16.0%) vs. females (9.9%) .

Comparative Analysis of Anti-AAV Antibodies

Antibody	Binding Region	Neutralization Strength	Clinical Utility
ADK9	3-fold axis	Strong	Exclusion biomarker for Zolgensma® trials
HL2374	Near 3-fold axis	Weak	Limited impact on spleen transduction
HL2368	2/5-fold wall	Non-neutralizing	No exclusionary effect

Therapeutic Implications

Gene Therapy Eligibility: Patients with anti-AAV9 titers ≥1:50 are excluded from onasemnogene abeparvovec (Zolgensma®) .
Engineering Solutions: Mutagenesis of AAV9 capsids (e.g., S454A) enables evasion of ADK9-like antibodies without compromising tropism .

Research Gaps and Future Directions

No peer-reviewed studies or regulatory filings reference "ADF9 Antibody." The closest analogous agent, ADK9, highlights:

Unmet Needs: Universal AAV9 vectors resistant to preexisting immunity.
Technical Challenges: Balancing epitope modification with capsid stability and transduction efficiency .

Product Specs

Buffer

Preservative: 0.03% Proclin 300
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

ADF9 antibody; At4g34970 antibody; M4E13.30Actin-depolymerizing factor 9 antibody; ADF-9 antibody; AtADF9 antibody

Target Names

ADF9

Uniprot No.

O49606

Target Background

Function

ADF9 exhibits a unique characteristic in that it does not display the typical F-actin depolymerizing activity observed in other proteins. Instead, it demonstrates a high capacity to stabilize and cross-link actin filaments, functioning as an actin bundling protein. This activity is most efficient under acidic conditions. Furthermore, ADF9 may play a crucial role in regulating the levels of histone modifications, specifically histone H3 lysine 4 trimethylation and H3 lysine 9 and 14 acetylation at the FLC locus.

Gene References Into Functions

ADF9 orchestrates both cytoplasmic and nuclear processes within the shoot apical meristem, playing a critical role in the regulation of multicellular development. [ADF9] PMID: 18830798

Database Links

KEGG: ath:AT4G34970

STRING: 3702.AT4G34970.1

UniGene: At.31455

Protein Families

Actin-binding proteins ADF family

Subcellular Location

Cytoplasm, cytoskeleton.

Q&A

Based on the analysis of available research documents and patent literature, here is a structured FAQ addressing key scientific considerations for working with ADF-related antibodies in academic research:

Basic Research Questions

How to validate antibody specificity for actin depolymerization studies?
- Perform parallel validation using:
  - Peptide blocking assays (pre-incubate antibody with immunogen peptide)
  - siRNA knockdown controls (compare staining intensity in ADF-depleted cells)
  - Cross-species reactivity tests using conserved epitope regions
- Recommended positive controls: NIH/3T3 fibroblasts (dynamic actin edges) or A-431 carcinoma cells

What experimental controls are critical for immunofluorescence quantification?

Essential controls table:

Control Type	Purpose	Example Implementation
Isotype	Non-specific binding	Mouse IgG1κ for monoclonal studies
Fixation Artifact	Preservation of epitopes	Compare methanol vs. PFA fixation
Permeabilization	Accessibility validation	Titrate Triton X-100 (0.1%-0.3%)

Advanced Research Challenges

How to resolve contradictory data in actin filament binding assays?
Methodological considerations:
- Calcium chelation controls (ADF vs. cofilin differential responses)
- Quantitative FRAP parameters:
  - 488 nm laser intensity ≤5% to avoid actin photobleaching artifacts
  - Frame interval: 15 sec for G-actin dynamics
What validation strategies exist for phospho-specific isoforms?
- Triple approach:
  1. Phosphatase treatment: Lambda PPase incubation + western blot
  2. Motility correlation: Boyden chamber assays with pADF/cofilin inhibitors
  3. Structural validation: Cryo-EM density mapping of antibody-epitope interfaces

Technical Optimization

How to design longitudinal studies with antibody stability constraints?
- Stability data from comparable antibodies :
  
  Storage Condition Signal Retention (6 months) Recommended Use
  -20°C (lyophilized) 98% ± 2.1 Quantitative WB
  4°C (liquid) 74% ± 5.6 Qualitative IF
  RT (lyophilized) 81% ± 3.8 Emergency stock
- Aliquot strategy: 50 μL/vial with 0.1% BSA carrier protein

Storage Condition	Signal Retention (6 months)	Recommended Use
-20°C (lyophilized)	98% ± 2.1	Quantitative WB
4°C (liquid)	74% ± 5.6	Qualitative IF
RT (lyophilized)	81% ± 3.8	Emergency stock

Emerging Research Applications

What novel delivery methods enhance antibody penetration in 3D models?
- Electroporation parameters for organoid systems:
  - Voltage: 100-120 V/cm (pulse duration 5 ms)
  - Validation metric: Antibody distribution depth via z-stack analysis
- In silico modeling approach:
  - Use Brownian dynamics simulations with antibody size (≈150 kDa) and matrix porosity

Data Interpretation Framework

How to distinguish ADF-specific effects from cofilin compensation?
- Experimental design matrix:
  
  Condition ADF Knockout Cofilin Inhibitor Expected Outcome
  1 Yes No Baseline phenotype
  2 No Yes Compensation threshold
  3 Yes Yes Synthetic lethality

Condition	ADF Knockout	Cofilin Inhibitor	Expected Outcome
1	Yes	No	Baseline phenotype
2	No	Yes	Compensation threshold
3	Yes	Yes	Synthetic lethality

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ADF9 Antibody