Antibody as a powerful tool for diagnosis and research, its quality problem has been paid close attention by researchers worldwide. In the 2003 Human Protein Atlas (HPA) program, researchers validated more than 9000 antibodies from 29 vendors using WB and IHC, including monoclonal and polyclonal antibodies, and found that 49% were completely ineffective, while only 7% of the antibodies proved to be fully potent and possessed pre- and post-experimental consistency. According to statistics, before 2016, the annual loss of funding for research due to the use of lack-of-validated antibodies was approximately US $800 million in the world. The "antibody crisis" created by the cross reactivity versus variability of antibodies continues to present each year with substantial loss of funding for research.
WB, IHC, peptide arrays, and protein arrays, and so on are the commonly used methods for antibody validation, but they all are not perfect with more or less disadvantages, such as time-consuming operation, inability to validate in high throughput, difficulty in quantification, and difficulty in standardization, which means that one antibody may perform well in one validation method but fail completely in the other. So traditional validation methods alone do not guarantee specificity in this antibody, and many antibody companies are actively searching for new alternatives to demonstrate the quality of their antibodies to the consumer.
In 2014, it was reported that a well-known antibody company began trying to employ siRNA technology to inhibit target protein expression for specificity verification by antibody products. So far, however, only a few dozen antibodies have been validated by siRNA across ten thousand products, and it is clearly to see the efficiency of this validation method (siRNA-mediated gene knockdown models) is not high.
Choosing an efficient and accurate method for antibody validation will not only guarantee antibody quality but also significantly improve the corporate competitiveness and brand image of antibody companies. Thus, a number of antibody companies are looking for better validation methods for antibodies. Abcam, the global antibody lead company, launched a series of antibodies validated using KO cells in 2019, meanwhile, also eliminated more than a thousand antibodies that failed in validation, creating a new standard of high-quality antibodies, and opening the "Golden" standard for antibody validation.
In this figure, Ubigene randomly selected 7 EGFR antibodies from different commercial brands on the market, and their specificities were verified using EGFR-KO MC3T3 cell lysates. Validation results showed that four antibodies performed good in specificity and did not show any bands in the KO lane; Three other EGFR antibodies were less specific and displayed smeared bands.Therefore, we can know there are still antibodies with poor specificity on the market, and Ubigene's KO cell lysate can precisely and efficiently verify the specificity of antibodies.
Validation of antibody specificity by KO cells, by virtue of their unique strengths, has become the "Golden" standard for antibody validation. The table below is a comparison of technical advantages and disadvantages of KO cell models and siRNA knockdown models:
| Validation methods | Advantages | Disadvantages |
|---|---|---|
| KO cell models (single-cell clones) | KO on DNA level, loss of expression of target protein, "Golden" standard for antibody validation Optimal for antibody specificity validation, a perfect negative control Applicable for large-scale bulk antibody screening | Some essential genes make it difficult to construct KO cell lines Relatively high cost, difficult to bulk production |
| （pool）KO cell models (cell pool) | KO on DNA level, down-regulation of target protein expression, applicable for antibody validation Strong, convincing negative control for antibody specificity validation Applicable for large-scale bulk antibody screening Applicable for large-scale bulk antibody screening | Some cell lethal, apoptotic genes make it difficult to construct KO cell lines Incomplete KO (polyclonal) Incomplete KO (polyclonal) |
| siRNA knockdown models | Gene knockdown on RNA level, down-regulation of target protein expression, can be used for antibody validation Can be used for antibody specificity validation, traditional negative control Applicable for large-scale bulk antibody screening | Some cell lethal, apoptotic genes make it difficult to construct knockdown cell lines Knockdown on RNA level, which means it is difficult to regulate the knockdown level of target protein expression Unstable siRNA efficiency, incomplete KO Non DNA level knockdown, less convincing |
As an enterprise focused on the field of gene-editing cells, Ubigene by virtue of the exclusive Red Cotton™ gene editing system and CRISPR-U™ patented technology of gene editing, has successfully developed UBI-001 near haploid cell line over 2 years, it enables us to stably batch provide high-quality KO cell lysates.
Adherent cells, fast proliferation, easy-to-transfect, strong colony formation ability
Near-haploid cell line, high KO effeciency, nearly 100%
With a team of experts with over 12 years of experience in gene-editing, Ubigene has so far successfully KO over 5000 genes in over 100 types of cell lines, and successfully established an exclusive KO Cell Bank and gRNA Plasmid Bank, covering 10 major research areas, 8 major signaling pathways, 40 diseases, and over 1300 genes. Currently, Ubigene's annual capacity of KO cell lysates is expected to be over ten thousand.
Ubigene's exclusive and intelligent data processing system - Red Cotton™ CRISPR gene-editing system enables to batch and automatically generate gene knockout strategies, which is the foundation for high-throughput production of KO cell lysates.
Ubigene exclusive developed CRISPR-U™ system, comparied with conventinal CRISPR/Cas9 technology, its cutting efficiency is 10-20 times higher, providing the technical guarantee for the production of high-quality KO cell lysates.
Ubigene's EZ-editor™ series products and IT tools can greatly simplify the preparation procedure of KO cell lysates, also further improve the cell transfection efficiency and single-cell clone formation rate, effectively scaling down the turnaround of services and production by at least 25%.
More than 600 types of KO cell lysates (monoclonal) are now available in Ubigene, as well as custom KO cell lysate services >>
| Product & Service | Turnaround | Quantity | Price | |
|---|---|---|---|---|
| KO cell lysates (clone) | In-stock | 1 week | 150ug/vial/gene | Starting from 320 USD |
| Custom service(UBI-001) | 6-8 weeks | 150ug/vial/gene | Please Inquire >> | |
Custom KO cell lysates (pool) service is now available in Ubigene, cutting effect verified by polyclonal sequencing and KO effect guaranteed >>
| Product & Service | Turnaround | Quantity | Price | |
|---|---|---|---|---|
| KO cell lysates (pool) | Custom service(UBI-001) | 4-6 weeks | 150ug/vial/gene | Please Inquire >> |
EZ-editor™ Series Products are the gene-editing innovation products innovatively developed by Ubigene, including 6 major product series of cell lines, culture medium, kits, gRNA plasmids, lentiviruses, KO cell lysates, comprehensively covering the experimental flow for gene-editing. Our products enable simplification of experimental flow and significant improvement of gene-editing efficiency. And the EZ-editor™ KO cell lysates will be the key words for a boom in antibody validation in the future!
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