Great bonds make Great Proteins.

Integrated services to take your molecule from gene sequence to potential drug candidate. Across multiple host systems - bacterial, yeast and mammalian.

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Expression Scouting

Production of recombinant proteins has led to the advancement of therapeutics, vaccines and other relevant industries. Need to purify proteins in an easy, cost-effective, scalable and reproducible manner is of primary importance. A decision to select the most appropriate expression platform is a challenge the industry faces at the onset of program, and to this need, Premas has developed our expression and purification screening platform. This is primarily to fill the void of technology where the scientists are forced to use same or similar systems to express simple as well difficult to express proteins.

Premas Biotech offers a a rapid expression optimization platform that can screen between 200 to more than 2000 data points across four host expression systems, namely bacteria, yeast, and mammalian cells. This has been extremely impactful in our studies involving the expression of “Difficult to Express Proteins” (DTEP). We have successfully used this platform to screen and obtain conditions for expression for proteins ranging from 2-3 to as high as 15-24 multi-span membrane proteins, which are either novel or have limited data available. Through our platform, we identify the key parameters that have been observed to play a pivotal role in optimising expression for DTEPs. The results produced are rapid and definite and can be further used for a much rational approach design for production of these proteins.

This strategy has been utilised to deep scout DTE proteins, and over 250 proteins which are membrane, or membrane bound, highly hydrophobic (>50% hydrophobicity) and high probability to aggregate. The use of the deep scout design has been successfully implemented to rapidly deliver data to enable correct decisions to be taken.

Premas has also utilised the same deep screen to develop highly effective strategies for obtaining high yielding combinations of cell line/strain, vectors, and expression conditions. We have achieved the typical range 1-6 gm/L in various expression systems.

Purification Screen

Protein purification is executed in scales from micrograms and milligrams in research laboratories to kilograms and tons in industrial sets. This mandates a significant amount of time and effort being invested in developing a robust, reproducible and cost-effective purification process.

Premas has developed and delivered using its purification screens, multiple proteins ranging from milligrams to kilograms in a timely manner. With difficult to express and purify proteins, this become even more imperative to implement to isolate and purify integral membrane proteins, unstable protein complexes, proteins expressed as insoluble aggregates, and proteins with a specific set of post-translational modifications. This, the design of the screen is of paramount importance.

Premas purification screen matrix allows for the simultaneous testing of multi-modal methods in a parallel manner to achieve the target profile of a protein

The salient points of the screen:
• rapid generates focused data
• simultaneous validation along with in silico analysis
• yields material for testing
• suggests processes and resins which are easily scalable, robust and leading to ideal COGs

Premas have delivered hundred’s of proteins using the purification screen.

Formulation Study

Difficult to express proteins like membrane associated, presence of high number of cysteine’s and free cysteine’s along with high number of proline residues pose a challenge in producing soluble expression and subsequent challenges in purification. The aggregation pathway is being studied thoroughly. A schematic diagram is mentioned below to understand the proteins aggregation pathway.

Difficult to express proteins are challenging to work with, due to their susceptibility to degradation and aggregation, so protein stability screening is an important step. The need for the early formulation study is to minimize the adverse effects in the mid or later stage of the development. It also impacts the cost of goods (COGs).

To cater to this need of our customers, Premas has developed a 48 well plate method to optimize a number of variables to maximize product stability under the recommended storage conditions. This study consists of screening of possible formulations, including buffers, additives, stabilizers, excipients, etc to stabilize the protein against physico-chemical degradation and aggregation over the desired shelf life. The factors which greatly influence protein stability in a solution are temperature, pH, buffer composition in terms of buffer, salt, metal-ions, co-solvents, preservatives, surfactants, protein concentration, freeze thaw cycles etc.