The Ultimate Array test for Validating Antibody Specificity
High specificity is the pre-requisite for any antibody used in diagnostic and therapeutic applications. Antibody cross-reactivity will create unexpected side effects of false diagnostic reports for clinicians. Research data from various groups have shown that some monoclonal antibodies on the market are not mono-specific. However, there was no commercial tool available to test the specificity of an antibody thoroughly.
OriGene Offers A Solution: UltraMAB Array Test
OriGene now provides UltraMAB Service, the ultimate array test to validate the antibody specificity. The UltraMAB Service leverages OriGene's proprietary technology platform, the High-Density Protein Microarray, to test the specificity of an antibody against the human proteome in a high-throughput manner. This platform can also be used to screen for the target(s) of an antibody or a small molecule.
What is High-Density Protein Microarray?
With a large collection of overexpression antigen standards, OriGene developed a high-density protein microarray chip for antibody evaluation at the proteome-wide level. This protein chip is spotted with over 10,000 overexpression proteins in duplicate on a single nitrocellulose coated glass slide, as well as various positive and negative controls. The array was manufactured as indicated in the flow chart below and tested with 1:500 dilution of OriGene's anti-DDK (FLAG) tag antibody followed by DyLight 649 conjugated goat anti-mouse IgG secondary antibody.
Specifications of the High-Density Protein Microarray
OriGene protein microarray slide specifications:
|Slide type:||White nitrocellulose glass slide|
|Slide NC pad dimension:||20mm x 60mm|
|Total subarrays:||48 (4 columns x 12 rows)|
|Subarray Size:||4200um x 4400 um|
|Subarray Dimensions:||21 rows (Vertical) x 22 columns (Horizontal)|
|Median Spot Diameter:||~120um|
|Spot Center to Center Spacing:||200um|
|Distances Between Subarrays:||100um|
|Replicates per Sample:||2|
Under current specifications of the 10K microarray chip, 10,728 samples were spotted in duplicates on a single nitrocellulose slide. These include 10,020 overexpression lysates and 708 positive and negative control samples. The premium 17K chip contains 16,242 overexpression lysates. Serial diluted mouse, human and rabbit IgG mixture as positive controls were spotted on three subarrays highlighted in redboxes. Subarrays highlighted in yellowboxes contain serial diluted HEK293T cell lysates as negative controls. All the subarrays contain standard positive and negative controls as shown in the figure above. The major protein family coverage is shown in the figure below.
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UltraMAB Array Test - Scope of Service
With the UltraMAB array service, OriGene helps you identify reactive proteins of your antibody and potential off-target effects to eliminate the cross-reactivity concerns. Please refer to the flow chart below for the details of this service
The current service offers our standard 10,000 or platinum 17,000 human protein array. The average timeline for array chip test and data analysis is 3 weeks upon receiving customer's antibody.
Examples of UltraMAB Array Tests
OriGene has applied the High-Density Protein Microarray to test the specificity of over 100 UltraMAB® and other commercial antibodies. Here we showcase two case studies of the comparison between UltraMAB and other antibodies with the microarray.
Case Study 1: HER2 antibodies for breast and gastric cancers - UMAB36 vs. 4B5
Case Study 2: ERCC1 antibodies for non-small cell lung cancer - 4F9 vs. 8F1
If you are interested in testing the specificity of your antibody, please contact firstname.lastname@example.org