Intracellular Non-Aggregate ScFv Screening Service

Creative Biolabs has established a novel strategy to screen the non-aggregate scFv under the cytoplasm environment. Through fusion with our novel Super™ signal protein, our scientists are able to validate whether the interest scFvs will aggregate when transfer into the cells, and contribute to the selection of functional intracellular scFvs or improve the quality of the scFv-based intrabody library.

Intracellular Screening of Non-Aggregate scFv via Super™ Signal Protein

Since the scFv aggregates in the cytosol of mammalian cells are always existed and leading to lower product bioactivity and yield, our platform has adopted an advanced strategy to eliminate aggregated-caused scFvs. To reach this goal, the interest scFvs will be fused with our Super™ signal protein. The non-aggregate fused scFvs will not play extra roles to affect the host cells. However, once the aggregation of scFvs occurred, the aggregate Super™ signal protein will stimulate the signal pathway of host cells and then result in significant phenotype change. In this way, the normal cells with non-aggregate intracellular scFvs can be screened. The sequence of non-aggregate scFvs will then be selected for further tests.

Improvement of scFv Libraries for Intrabody Selection

In terms of the similar concept, our Super™ signal protein can also contribute to improving the quality of scFv libraries used for intrabody screening. The high-quality scFv libraries are always ideal source for the hit identification, especially for the intrabody discovery and development. Recently, accumulated research data have indicated a vicious phenomenon of scFv aggregation from a scFv library expressed in the cytosol of mammalian cells, which causes lower antibody bioactivity and product output. While based on our Super™ signal protein technique, it is possible to generate a Super™ signal protein fused-scFv pool and eliminate most of the aggregate binders through the intracellular selection. A high-quality scFv library contains intracellular non-aggregate scFvs can be constructed as a novel source of intrabody development. This strategy is a potential solution to reveal the stable and soluble scFvs which could be the most desirable candidates for intrabody selection.

Fig. 1 Screening of Non-Aggregate scFv Fusion with Super™ Signal Protein.

Subsequently, our scientists are able to perform our optimized intrabody screening approaches (e.g. IAC technology, bacterial two-hybrid technology, and mammalian two-hybrid technology) to identify the suitable intrabody for the interest targets. It is also a potential option to employ our specific positioning intrabody development technology to achieve specific intracellular localization of the select intrabodies. Meanwhile, our scientists can perform multiple strict validation tests on multiple parameters based on in silico analysis to achieve the best specificity and highest bioactivity.

At Creative Biolabs, we are keeping alert for any novel and advanced technology progress to assist our clients’ valuable projects. Our scientists are dedicated to overcoming almost every bottleneck in the discovery and development of specific intrabodies. If you are interested in developing novel intrabodies, please do not hesitate to contact us for detailed information.

All of our antibody products and services can only be used for preclinical research studies. Do not use them on humans.