ACROBiosystems – Full length CD20 with Nanodisc / DDM|Tokyo Future Style is a bio-tech trading company

FACS Verified and Protocol Offered

The new released article Structure of CD20 in complex with the therapeutic monoclonal antibody RTX published on Science by Genentech team highlights the importance of having intact CD20 for antibody drug development. The team have discovered several RTX mutants that, despite maintaining cell-binding activity comparable to that of OBZ, and largely unaffected binding to the primary ECL2 epitope, are incapable of eliciting CDC. This confirms that the secondary ECL1/2 epitope and Fab:Fab interface contribute to the unique binding properties and high CDC activity of RTX.

Quotation: Lionel Rougé, et al. Structure of CD20 in complex with the therapeutic monoclonal antibody RTX. Science. 2020

To help CD20 antibody drug development, the HEK293 expressed full length CD20 from ACROBiosystems has been very attractive to customers.

BackgroundDifferent TypesProduct ListFeatures

Background

The 61st annual meeting of the American society of hematology (ASH) in 2019 presented bi-specific antibodies targeting CD20 on the surface of B cells and CD3 on the surface of T cells, such as CD20- TCB, with preliminary resultsof clinical studies showing acceptable safety and promising efficacy of this pair of specific antibodies.

CD20 is a hot target for the treatment of lymphoma, leukemia and some autoimmune diseases. However, due to low expression titer and the unstable bioactivity, it is so difficult to find an active CD20 protein on the market for antibody drug development and the CD20 CAR expression.

To address these issues, ACROBiosystems has developed the highly active full length multi-transmembrane CD20 proteins, expressed by HEK293 cells. The active full length CD20 proteins carry both big and small ECD loops, and bind RTX and OFA with high binding affinity in ELISA and SPR assay. We have different types of CD20 proteins include unconjugated protein, biotinylated proteins with DDM and Nanodisc, which are suitable for immunization, ELISA, SPR, BLI and FACS.

Related Page: Introduction of ACROBiosystems
Related Page: Bispecific antibody targets

Different Types of CD20 Proteins

Protein with DDM.

DDM can disintegrate the lipid membrane to release the membrane protein and provide a hydrophobic environment, maintain and protect the hydrophobic transmembrane structure of membrane proteins. But it is not recommended for cell experiments because it can destroy the structure of the cell membrane.

Protein with Nanodisc.

Nanodisc is a non-covalent structure of phospholipid bilayer and membrane scaffold protein. Membrane proteins can be integrated into nanodiscs to maintain their biological activity, CD20 protein with nanodisc was recommended for cell experiments.

Product List

ACRO’s new product CD20 proteins have a complete and natural conformation and shown a high sensitivity without unspecific binding background (FACS verified). Protocols are provided for free. You can click the following link to request. Ask for Tokyo Future Style, Inc.

Cat. No.	Product Description
CD0-H52H1	Human CD20 / MS4A1 Full Length Protein, His Tag (Nanodisc) (HEK293)
CD0-H52H3	Human CD20 / MS4A1 Full Length Protein, His Tag (HEK293) (SPR verified)
CD0-H82E3	Biotinylated Human CD20 / MS4A1 Full Length Protein, His,Avitag™ (Nanodisc) (HEK293)
CD0-H82E5	Biotinylated Human CD20 / MS4A1 Full Length Protein, His,Avitag™ (HEK293)

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Product Features

Full length CD20 protein has a complete conformation with both small and big loops.

Full length CD20 protein has better bioactivity

A) HEK293 Cell (Full Length)		B) E.coli (Extracellular)
Immobilized RTX at 2 μg/mL (100 μL/well) can bind Human CD20 Full Length, His Tag, HEK293 (SPR verified) (Cat. No. CD0-H52H3) with a linear range of 0.4-3 ng/mL (in presence of DDM and CHS).		Immobilized RTX at 5 μg/mL (100 μL/well) can bind Human CD20, TrxA Tag, His Tag (E.coli) with a linear range of 78-2500 ng/mL.

The bioactivity of full length CD20 protein was verified by FACS/ELISA/ SPR and the protocols are offered.

FACS

Cat. No. CD0-H82E3 (HEK293 expression)	Cat. No. CD0-H52H1 (HEK293 expression)
FACS Analysis of anti-CD20 CAR expression	FACS Analysis of anti-CD20 CAR expression
2e5 of CD20-CAR-293 cells transfected with anti-CD20-scFv were stained with 100 μL of 3 μg/mL of Biotinylated Human CD20 Full Length, His,Avitag (Cat. No. CD0-H82E3) and negative control antibody respectively, washed and then followed by PE-SA and analyzed with FACS.	2e5 of CD20-CAR-293 cells transfected with anti-CD20-scFv were stained with 100 μL of 3 μg/mL of Human CD20 Full Length, His Tag (Cat. No. CD0-H52H1) and negative control antibody respectively, washed and then followed by PE anti-His antibody and analyzed with FACS.

ELISA

Cat. No. CD0-H82E3 (HEK293 expression)	Cat. No. CD0-H52H1 (HEK293 expression)
Immobilized RTX at 5 μg/mL (100 μL/well) can bind Biotinylated Human CD20 Full Length, His,Avitag (Cat. No. CD0-H82E3) with a linear range of 0.008-0.125 μg/mL	Immobilized RTX at 2 μg/mL (100 μL/well) can bind Human CD20 Full Length, His Tag (Cat. No. CD0-H52H1) with a linear range of 1-63 ng/mL

Cat. No.CD0-H82E5 (HEK293 expression)	Cat. No.CD0-H82E5 (HEK293 expression)
Immobilized RTX at 2 μg/mL (100 μL/well) can bind Biotinylated Human CD20 Full Length, His,Avitag (HEK293) (Cat. No. CD0-H82E5) with a linear range of 4-63 ng/mL (in presence of DDM and CHS).	Immobilized OFA at 2 μg/mL (100 μL/well) can bind Biotinylated Human CD20 Full Length, His,Avitag (HEK293) (Cat. No. CD0-H82E5) with a linear range of 4-63 ng/mL (in presence of DDM and CHS).

Cat. No. CD0-H52H3 (HEK293 expression)	Cat. No. CD0-H52H3 (HEK293 expression)
Immobilized RTX at 2 μg/mL (100 μL/well) can bind Human CD20 Full Length, His Tag, HEK293 (SPR verified) (Cat. No. CD0-H52H3) with a linear range of 0.4-3 ng/mL (in presence of DDM and CHS).	Immobilized OFA at 2 μg/mL (100 μL/well) can bind Human CD20 Full Length, His Tag, HEK293 (SPR verified) (Cat. No. CD0-H52H3) with a linear range of 0.4-6 ng/mL (in presence of DDM and CHS).

SPR

Cat. No. CD0-H82E5 (HEK293 expression)	Cat. No. CD0-H52H3 (HEK293 expression)
Biotinylated Human CD20, His,Avitag (HEK293) (Cat. No. CD0-H82E5) captured on Biotin CAP-Series S Sensor Chip can bind RTX with an affinity constant of 1.73 nM as determined in a SPR assay (in presence of DDM and CHS) (Biacore T200).	Human CD20 Full Length, His Tag, HEK293 (SPR verified) (Cat. No. CD0-H52H3) captured on CM5 chip via Anti-human IgG Fc antibodies surface can bind RTX with an affinity constant of 6.21 nM as determined in a SPR assay (in presence of DDM and CHS) (Biacore 8K).

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