Quick adme quote
Applying understanding of protein structure
to support drug discovery progress
for Apini Therapeutics
Delivering functional, dimeric NAMPT protein
to support downstream assays
Project overview
The challenge
- The use of recombinant NAMPT protein from a number of providers did not provide expected results in biophysical and phenotypic screens.
- This would significantly delay program progress.
Our solution
- Applied structural biology and protein engineering expertise to redesign the construct.
- Delivered active, high-purity, low-endotoxin protein.
Our impact
Enabled consistent biophysical assay read-outs and supported phenotypic assay development, contributing to the progression of the client’s drug discovery program.
“The Protein Science department at Concept Life Sciences succeeded in preparing high quality and functional protein by identifying a foundational problem with our protein supply. The team then successfully produced high quality, active and low-endotoxin protein.
This directly enabled significant and consistent biochemical, biophysical and phenotypic read-outs, allowing our program to progress.
We were very impressed with this work and are grateful for their efforts!”
Sam Butterworth, CSO, Apini Therapeutics Ltd.
Read on for a detailed case study below
The challenge
A client approached us after working with recombinant NAMPT protein from a range of sources. These proteins showed variable profiles in biophysical assays and failed to elicit a consistent cytokine response in downstream phenotypic assays. As a result, progress on their program had stalled.
Our approach
- In-depth structural biology expertise - Enabled rapid diagnosis of a root cause of protein issues.
- Understanding of downstream requirements - Informed construct design to ensure the protein was active and fit for purpose.
- Tailored purification strategy – Delivered high-purity, low-endotoxin, active protein suitable for functional assays.
Our solution
We began by performing a detailed structural assessment of NAMPT, focusing on its dimeric form. This review revealed that protein from previous suppliers had a tag positioned adjacent to the dimer interface; a critical region required for correct oligomerization. This tag placement potentially disrupted dimer formation, thereby impacting the protein’s biological function.
To address this, we engineered a new expression construct designed specifically to preserve the dimer interface. By repositioning the affinity tag away from this region, we enabled correct dimer assembly.
We then developed a tailored purification workflow to produce recombinant NAMPT at scale. The protocol was optimized to deliver protein with:
- High purity (>95%) suitable for crystallography (Figure1)
- Verified dimeric conformation (Figure 2)
- Functional activity in relevant downstream cellular assays (Figure 3)
- Low endotoxin levels, supporting use in phenotypic cell-based systems
This end-to-end approach restored NAMPT activity and ensured the protein was fit for downstream applications.
The result
We delivered high-quality, functional protein that met all the client’s requirements:
- Active and dimeric
- Crystallographic-grade purity (>95%)
- Low endotoxin levels
- Unblocked downstream studies and enabled progression of the client’s drug discovery program
Let's talk about what we can do for your next project