As the largest human membrane protein family responsible for functions such as sensory regulations and cellular responses, GPCRs are considered one of the most valuable drug targets in the pharmaceutical industry. Abnova has developed a robust, reliable, and cost-effective high throughput platform of proteoliposome reconstitution system and monoclonal antibody (mAb) production targeting GPCRs to overcome the hindrance of lacking suitable antigens for producing high-quality mAbs.

By optimizing cell-free protein synthesis in the presence of liposomes and a purification process via discontinuous sucrose density gradient centrifugation, soluble reconstituted GPCR proteoliposome antigens are synthesized. Abnova’s exclusive techniques in manufacturing proprietary bioreagents allow us to not only produce GPCR proteoliposomes, amenable to functional analysis but also generate customized mAbs with sensitivity, specificity, and wide dynamic range. A high successful rate on customized GPCR mAbs has also expanded our field to deliver mAbs targeting various membrane proteins as well.

Related FAQ

Technology Comparison for Proteoliposome Expressions

 

Conventional Expression System

Bilayer / Dialysis Technique

Bilayer Automation System

Stability

Low Intermediate High

Solubility

Low Intermediate

High

Yield

Low Low

High

Versatility

Low Intermediate

High

Throughput Low Low

High

 

Six Categories and 300+ GPCR Proteoliposomes

 

Catalog Products

Made to Order

Adhesion Class

6 6

Class A

129

62

Class B

6

3

Class C

6

6

Class Frizzled 3

0

Others 9

62

 

 

Demonstrations

Bioactivity Profile on GPCR Protein Synthesized by Bilayer Automation System

 GPCR-SPR.jpg (16 KB)

 

Surface Plasmon Resonance (SPR) Analysis of Wild-type DRD1

The ligand-binding activity of DRD1 was determined using SPR in which dopamine and histamine were immobilized onto the measuring cell and reference cell at the same level. Wildtype DRD1 specifically bound to dopamine at KD value of 0.7X10-6 M.

 

Characterization of Abnova's Mab Against GPCR Proteins

 

 

Affinity Analysis of anti-DRD1 Mouse Mabs

The affinity of 36 mice Mabs was evaluated using a Scatchard plot with ELISA. The Kd values of mouse Mab ranged from 10-7 M to 10-10 M, with half of the population showing high sensitivity toward DRD1.

 

 

 

Specificity Analysis of GPCR Mouse Mabs

Mouse Mabs against 3 different GPCRs, including GHSR (Class A), PTGER1 (Class A), and T1R1 (Class C), were evaluated using ELISA. All anti-GPCR mAbs showed specific interaction with its intended target antigen.

 


References

Takeda, H., et.al. (2015). Scientific Reports. DOI: 10.1038/srep11333
Goren, M.A. and Fox, B.G. (2008). Protein Expression and Purification. DOI:10.1016/j.pep.2008.08.002
Nozawa, A., et. al. (2016). Plant Cell Physiology. DOI:10.1093/pcp/pcm150

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