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Simple Western

Simple Western™ is a CE platform that pairs protein size or charge separation with a quantitative immunoassay readout and represents the next-generation gold standard for quantitative, reproducible, and highly specific analytical-grade protein expression potency assays. Plus, Simple Western is a multi-attribute method used in cell and gene therapy process development for quantitative CQA monitoring of purity, titer, empty/full capsid content, and capsid protein ratio.

  • Quantitation. Quantitative protein measurements with a 4-log dynamic range.
  • Sensitivity. Conserve your precious AAV sample with CQA analysis, including purity assays that are more sensitive than SYPRO Ruby.
  • Reproducibility. Eliminate the variation of traditional Western blots -CVs below 15-20%.
  • High Throughput. Up to 96 data points in a single overnight run, all in a fully automated fashion.
  • Method Transferability. Easily transfer between sites or from sponsors to CROs. 21 CFR Part 11-compliant and GMP-ready.
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Gene Therapy

Simple Western relative protein expression potency data analysis

The New Gold Standard for Transgene Expression Assays

Get the Flexibility of Western Blots with the Analytical-Grade Quantitation of ELISA

By pairing the flexibility and specificity of Western blots with the analytical-grade quantitation of ELISA, Simple Western is ideal for making specific measurements in complex sample types like tissue homogenates and analyzing intracellular protein expression without matrix effects that can confound ELISA read-outs. As a platform technology that needs only a single validated antibody and offers rapid 3-hour runtimes, Simple Western lowers the custom assay development hurdles of ELISA and accelerates drug development from pre-clinical phases and scaling to GMP. In this Application Note, we demonstrate a Simple Western relative potency assay with parallel line analysis (PLA) for AAV-mediated gene delivery in cultured human cells.

One Platform, Many Assays to Support Your Process and Product

3 Hours to Results, Just 3 µL of Sample

AAV Identity and Purity on Simple Western

Identity and specificity provided by an anti-VP1/2/3 antibody shown in green. Purity provided by total protein stain in light grey.

HEK293 HCP Total Protein detection on Simple Western

Identity and specificity provided by an anti-HEK HCP antibody shown in the top. Purity provided by total protein stain shown in the bottom panel. 

Dystrophin protein expression analysis by Simple Western

The guidance for industry submitted to the FDA recommends Simple Western for reproducible dystrophin quantification.

AAV capsid content ratio

Simultaneous detection of VP1/2/3 and DNA content of AAV particles enables quantification of the % full AAVs in a sample.

AAV Capsid Protein Ratio

VP1:2:3 capsid protein ratios can be measured to quantify VP molar ratios for optimization of infectivity and potency of AAV gene therapy products.

p24 Lentivirus titer quantification with Simple Western

Titer assays with linearity and high-sensitivity in an automated workflow optimized for AAV and lentiviral samples.

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Immune Cell Therapy

When it comes to viral vector characterization for cell and gene therapies, Simple Western isn't just limited to AAVs. With Jess, you get multi-attribute analysis for lentivirus as well, including titer, purity, capsid content, stability, transduction analysis, and more.

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Regenerative Medicine

Get unrivaled multiplex flexibility, sensitivity, and reproducibility for accurate protein expression measurements with automation that can scale in industrial stem cell differentiation and manufacturing workflows for regenerative medicine.

Single-Cell Analysis

Single-Cell Westerns on Milo for Heterogeneity Analysis

Single-Cell Westerns on Milo measure protein expression in thousands of single cells per run. Their high sensitivity combined with the ability to use conventional Western blotting antibodies makes them particularly well-suited for quantifying gene therapy efficacy and tracking stem cell differentiation.


    Cell culture
    HSC Hematopoietic Stem Cell Supplements, Media, and Reagents

    Milo's Single-Cell Westerns are used in clinical trials aimed at advancing gene therapies for Sickle Cell Disease and other hemoglobinopathies. Milo resolves hemoglobin subunits in single red blood cells, enabling researchers to track the efficacy of sickle cell gene therapies.

    Image of human iPSCs stained with antibodies to OCT3 and F-actin and counterstained with DAPI

    Single-Cell Western analysis can track phenotypic marker heterogeneity over time and monitor the relative proportion of cell subsets during culture differentiation of iPSC-derived cardiomyocytes. 

    Differentiated neurons stained for the expression of beta-III-tubulin and cell nuclei.

    Milo can identify and quantify neural subtypes in a heterogeneous neural sample, and monitor the differentiation of iPSCs into neurons, astrocytes, and oligodendrocytes using our research-grade or GMP differentiation reagents.

    Webinars in Cell and Gene Therapy Research

    Testing for Lentivirus and AAVs

    Get rapid assessment of viral titer and purity analysis for both lentiviral vectors and AAVs. Plus, get insight into AAV VP1:VP2:VP3 capsid protein ratio and AAV empty/full capsid content ratio.


    We introduce PROGEN's recombinant VP standards used with Simple Western automated capillary electrophoresis in fit-for-purpose solutions for viral titer and VP capsid protein ratio measurements.

    Customer Success Stories