MABSOL® BIOTINYLATED PROTEIN COLLECTION

                   
Avitag^TM Biotin-labeled Proteins        Chemically Biotinylated Proteins
@ PrecisionAvi Series　　　　　　    @ UltraLys Series

INTRODUCTION

The interaction between biotin and avidin/streptavidin (SA) is incredibly strong and specific, making it an excellent tool in protein biochemistry. However, generation of high quality biotinylated proteins presents a major challenge. Researchers often run into technical issues including insufficient labeling, high batch-to-batch variation, and unpredictable performance. In addition, the choice of biotinylation methods also significantly affects the ultimate outcome. Each target protein (assay) has its own unique biochemical characteristics, and therefore requires customized solution.

These confounding factors make it difficult for individual labs to make and use biotin conjugates, while at the same time few products are available commercially. To fill in this gap, Acrobiosystems has developed an exclusive collection of pre-labeled and experimentally validated biotinylated recombinant proteins. The Mabsol® biotinylated protein collection includes more than a hundred commonly studied drug targets and biomarker proteins. ACRO use high quality authentic human proteins as starting materials. ACRO thoroughly go through every option to achieve the highest bioactivity and detection sensitivity. ACRO also apply the most stringent quality control measurement possible to ensure minimal batch-to-batch variations.

Mabsol^® biotinylated protein collection includes two unique and complimentary product series, the PrecisionAvi series built upon the Avitag^TM technology, and the UltraLys series produced using the in-house developed chemical labeling method. These products are made with every attention to details.

PRODUCTS

PrecisionAvi Series

An exclusive collection of ready-to-use Avitag^TM biotinylated proteins

The products in this series are exclusively produced using the Avitag^TM technology. Briefly, a unique 15 amino acid peptide, the Avi tag, is introduced into the recombinant protein during expression vector construction. The single lysine residue in the Avi tag is enzymatically biotinylated by the E. Coli biotin ligase BirA.

This single-point enzymatic labeling technique brings many advantages for commonly used binding assays. The biotinylation happens on the lysine residue of Avi tag, and therefore does NOT interfere with the target protein’s natural binding activities. In addition, when immobilized on an avidin-coated surface, the protein orientation is uniform because the position of the Avi tag in the protein is precisely controlled.

PrecisionAvi Product List (Please refer to the Complete Product List (EXCEL)) for more information）

PCSK9
PC9-H82E7	Biotinylated Human PCSK9, Avi Tag (Avitag™)
PCY-H82E7	Biotinylated Human PCSK9 (D374Y), Avi Tag (Avitag™)
PC9-M82E1	Biotinylated Mouse PCSK9, Avi Tag (Avitag™)
PD1
PD1-M82F4	Biotinylated Mouse PD-1 / PDCD1, Fc Tag, Avi Tag (Avitag™)
PD1-H82F2	Biotinylated Human PD-1 / PDCD1, His Tag & Fc Tag, Avi Tag (Avitag™)
PD1-H82E4	Biotinylated Human PD-1 / PDCD1 (recommended for biopanning), Avi Tag (Avitag™)
PDL1
PD1-H82F3	Biotinylated Human PD-L1 / B7-H1, His Tag & Fc Tag, Avi Tag (Avitag™)
PD1-M82F5	Biotinylated Mouse PD-L1 / B7-H1, Fc Tag, Avi Tag (Avitag™)
PD1-H82E5	Biotinylated Human PD-L1 / B7-H1 (recommended for biopanning), Avi Tag (Avitag™)
PDL2
PD2-H82E8	Biotinylated Human PD-L2 / B7-DC Protein (recommended for biopanning), Avi Tag (Avitag™)
PD2-H82F6	Biotinylated Human PD-L2 / B7-DC, Fc Tag, Avi Tag (Avitag™)
FcRn
FCM-M82W6	Biotinylated Mouse FcRn / FCGRT & B2M, Avi Tag (Avitag™)
FCM-H82W4	Biotinylated Human FcRn / FCGRT & B2M, Avi Tag (Avitag™)
FCM-C82W5	Biotinylated Cynomolgus / Rhesus macaque FcRn / FCGRT & B2M, Avi Tag (Avitag™)
FCM-R82W7	Biotinylated Rat FcRn / FCGRT & B2M, Avi Tag (Avitag™)
EGFR
EGR-H82E7	Biotinylated Human EGF R, Avi Tag (Avitag™)
TNF-alpha
TNA-H82E3	Biotinylated Human TNF-alpha (HPLC-verified), Avi Tag (Avitag™)
TNA-M82E9	Biotinylated Mouse TNF-alpha (HPLC-verified), Avi Tag (Avitag™)
CTLA-4
CT4-C82E5	Biotinylated Cynomolgus CTLA-4, Avi Tag (Avitag™)
CT4-H82E4	Biotinylated Human CTLA-4, Avi Tag (Avitag™)
CT4-H82F3	Biotinylated Human CTLA-4, Fc Tag, Avi Tag (Avitag™)
OX40
OX0-H82F7	Biotinylated Human OX40, Fc Tag, Avi Tag (Avitag™)
VEGF
VE1-H82E7	Biotinylated Human VEGF121, Avi Tag (Avitag™)
CD40
CD0-H82F2	Biotinylated Human CD40 / TNFRSF5, His Tag & Fc Tag, Avi Tag (Avitag™)
CD47
CD7-H82F6	Biotinylated Human CD47, Fc Tag, Avi Tag (Avitag™)
IgG Fc
IG1-H82E2	Biotinylated Human IgG1 Fc, Avi Tag (Avitag™)
IGA-M8210	Biotinylated Mouse IgG2a Fc, Avi Tag (Avitag™)
TIGIT
TIT-H82F1	Biotinylated Human TIGIT Protein, Fc Tag, Avi Tag (Avitag™)
TIT-H82E5	Biotinylated Human TIGIT Protein, Avi Tag (Avitag™)
VISTA
B75-H82E1	Biotinylated Human B7-H5 / Gi24 / VISTA, Avi Tag (Avitag™)
HVEM / TNFRSF14
HV4-H82F1	Biotinylated Human HVEM / TNFRSF14, Fc Tag, Avi Tag (Avitag™)
Fc gamma RI/CD64
CD4-M52E8	Biotinylated Mouse Fc gamma RI / CD64, Avi Tag (Avitag™)
IGF-II
IG2-H82F9	Biotinylated Human IGF-II, Fc Tag, Avi Tag (Avitag™)
EGF R (vIII)
EGR-H82E0	Biotinylated Human EGF R (vIII), Avi Tag (Avitag™)
Fc gamma RIIA/CD32a
CDA-H82E7	Biotinylated Human Fc gamma RIIA / CD32a (R131), Avi Tag (Avitag™)
CDA-H82E6	Biotinylated Human Fc gamma RIIA / CD32a (H131), Avi Tag (Avitag™)
Fc gamma RIIB/CD32b
CDB-H82E0	Biotinylated Human Fc gamma RIIB / CD32b, Avi Tag (Avitag™)
CDB-M82E8	Biotinylated Mouse Fc gamma RIIB / CD32b, Avi Tag (Avitag™)
Fc gamma RIIIA/CD16a
CDA-H82E8	Biotinylated Human Fc gamma RIIIA / CD16a (F158), Avi Tag (Avitag™)
CDA-H82E9	Biotinylated Human Fc gamma RIIIA / CD16a (V158), Avi Tag (Avitag™)
Fc gamma RIIIB/CD16b
CDB-H82E1	Biotinylated Human Fc gamma RIIIB / CD16b, Avi Tag (Avitag™)

The following products are available for pre-order, please contact us for more information.

4-1BB	CD30	CD33	OX40	CD40/TNFRSF5	CD47
CD80/B7-1	FOLR1	LDLR	SIRPA/CD172a	TIM3/HAVCR2/KIM3	BACE
CD147	CD4	B7H4	CD38	cMet	CSF1R
EPCAM	ErbB3	IL-5	VEGFR2	CD22/SIGLEC2	INSR
CD86	B7-H5	TNFR1	GCSFR	CEACAM1	IL2R
IL6R	IL5R	CD319	TRAILR1	TRAILR2	TRAILR4
IL-10	IL-1a	IL-4	IL-6	CRP/C-Reactive	VEGFR1

※Unlabeled avi-tagged protein is available upon request.。

UltraLys Series

A unique series of chemically labeled biotinylated proteins with ultra sensitivity

The products in this series are produced using our in-house developed chemical labeling approach. The primary amines in the side chains of lysine residues and the N-terminus of protein are conjugated with biotins.

Chemical labeling usually results in multiple biotin attachment on a single protein molecule, which could potentially lead to higher detection sensitivity.

UltraLys Series Product List (Please refer to the Complete Product List (EXCEL)) for more information）

PD1
PD1-M8259	Biotinylated Mouse PD-1 / PDCD1, Fc Tag, ultra sensitivity (primary amine labeling)
CD19
CD9-H8259	Biotinylated Human CD19, Fc Tag, ultra sensitivity (primary amine labeling)
Her2/ErbB2
HE2-H822R	Biotinylated Human Her2 / ErbB2, ultra sensitivity (primary amine labeling, long spacer)
Her3/ErbB3
ER3-H8223	Biotinylated Human ErbB3 / Her3, ultra sensitivity (primary amine labeling)
VEGF165
VE5-H8210	Biotinylated Human VEGF165, epitope tag free, ultra sensitivity (primary amine labeling)
FcRn/FCGRT&B2M
FCM-H8286	Biotinylated Human FcRn / FCGRT & B2M, ultra sensitivity (primary amine labeling)
TNF-alpha
TNA-H8211	Biotinylated Human TNF-alpha, epitope tag free, ultra sensitivity (primary amine labeling)
TNA-H821R	Biotinylated Human TNF-alpha, epitope tag free, ultra sensitivity (primary amine labeling, long spacer)
B7-H4
B74-H8222	Biotinylated Human B7-H4, ultra sensitivity (primary amine labeling)
Siglec-3/CD33
CD3-H8226	Biotinylated Human Siglec-3 / CD33, ultra sensitivity (primary amine labeling)
FGF
BFF-H8117	Biotinylated Human FGF basic, epitope tag free, ultra sensitivity (primary amine labeling)
4-1BB/TNFRSF9
41B-H8258	Biotinylated Human 4-1BB / TNFRSF9, Fc Tag, ultra sensitivity (primary amine labeling)
CD3E&CD3D
CDD-H82W0	Biotinylated Human CD3E & CD3D, ultra sensitivity (primary amine labeling)
CD3 epsilon
CDE-H8223	Biotinylated Human CD3 epsilon, ultra sensitivity (primary amine labeling)
EphB4
EP4-H8229	Biotinylated Human EphB4, ultra sensitivity (primary amine labeling)
EpCAM/TROP1
EPM-H8223	Biotinylated Human EpCAM / TROP1, ultra sensitivity (primary amine labeling)
EPM-H8254	Biotinylated Human EpCAM / TROP1, Fc Tag, ultra sensitivity (primary amine labeling)
GM-CSF
GMF-H8214	Biotinylated Human GM-CSF, epitope tag free, ultra sensitivity (primary amine labeling)
GPA33
GP3-H8224	Biotinylated Human GPA33, ultra sensitivity (primary amine labeling)
TFPI
TFI-H8226	Biotinylated Human TFPI, ultra sensitivity (primary amine labeling)
Protein L
RPL-P814R	Biotinylated Recombinant Protein L, ultra sensitivity (primary amine labeling, long spacer)
IL-6
IL6-H8218	Biotinylated Human IL-6, epitope tag free, ultra sensitivity (primary amine labeling)
IL-1 alpha
ILA-H8213	Biotinylated Human IL-1 alpha / IL-1F1, epitope tag free, ultra sensitivity (primary amine labeling)
Mesothelin
MSN-H8223	Biotinylated Human Mesothelin / MSLN (296-580), ultra sensitivity (primary amine labeling)
MSN-H826x	Biotinylated Human Mesothelin / MSLN (296-580), Fc Tag, ultra sensitivity (primary amine labeling)
SCF/KITLG
SCF-M8228	Biotinylated Mouse SCF / KITLG, ultra sensitivity (primary amine labeling)
SOST/Sclerostin
SOT-H8245	Biotinylated Human SOST / Sclerostin, ultra sensitivity (primary amine labeling)
Transferrin R/CD71
TFR-H8243	Biotinylated Human Transferrin R / CD71, ultra sensitivity (primary amine labeling)
CX3CL1
CX1-H8221	Biotinylated Human CX3CL1, ultra sensitivity (primary amine labeling)

 

KEY FEATURES

High Quality Authentic Human Proteins

The production of our recombinant proteins including the biotinylated proteins is carried out using our proprietary HEKMax® expression platform. As expression hosts, the human HEK293 cells have a variety of advantages compared to other cell types as summarized in the table below. Most importantly, the proteins retain their authentic post-translational processing, which often translates into high bioactivity and stability.

Expression Systems	Folding	Phosphorylation	Proteolytic Processing	Glycosylation
E.Coli	+	N/A	N/A	N/A
Insect Cell	++	++	+	Poor
Plant Cell	+++	++	++	Poor
CHO Cell	++++	+++	+++	Non-Human Like
Human Cell	+++++	+++++	+++++	Human Authentic

 

High Bioactivity & Detection Sensitivity

The bioactivity of biotinylated proteins is determined both by the structure of the protein itself, and by the way how biotinylation is performed. For every single protein, ACRO test multiple options of tags and biotinylation methods, and evaluate the products in a variety of binding assays. Only those with the best performance are selected for production. Figure 1 is an example of our internal evaluation experiments:

 

PD1-Fc(N297A), enzymatically labeled, long spacer
PD1-Fc, chemically labeled, long spacer
PD1-Fc(N297A), chemically labeled, long spacer
 PD1-Fc(N297A), chemically labeled
 PD1-Fc, chemically labeled

 

 

 

 

Fig. 1 Binding activity of different forms of biotinylated PD1 evaluated in a functional ELISA against rhPD-L1 (Cat # PD1-H5258).

Our biotinylated proteins demonstrate high bioactivity and superb detection sensitivity. In the following Fig. 2, Biotinylated FcRn / FCGRT & B2M (Cat.# FCM-H82W4) coupled to SA coated sensor chip can bind Herceptin with an affinity constant of 2.24 μM as determined in a SPR assay. And in Fig. 3, when used in a functional ELISA, the biotinylated VEGF165 (Cat# VE5-H8210) can bind to immobilized rhVEGFR2 with a EC50 of 0.5-5 ng/mL.

Fig. 2 Biotinylated Human FcRn / FCGRT & B2M, Avi Tag (Avitag™) (Cat.# FCM-H82W4) coupled to SA coated sensor chip can bind Herceptin with an affinity constant of 2.24 μM as determined in a SPR assay (Biacore 2000). The data is generally provided by Biaffin GmbH &Co KG, Germany.

Fig. 3 The ELISA assay result of the biotinylated VEGF165 (Cat# VE5-H8210) which bind to immobilized rhVEGFR2.

Low Batch-to-Batch Variation

ACRO routinely apply rigorous quality control measures to ensure consistent performance of our products. Newly produced products are subjected to side-by-side comparison with our internal standard in a variety of assays. Only those within an acceptable margin of difference are allowed to be released. As demonstrated in Fig. 4, both EGFR-biotin batches show very similar KD values in a range between 104-112nM and are indistinguishable within the precision of the assay.
 
Fig. 4 Two lots of EGFR-biotin (Cat# EGR-H82E7) were immobilized on Sensor Chip SA in an SPR analysis to determine their binding affinity against EGF. Gradient concentration of EGF (ACRO’s product, EGF Cat# EGF-H4114  ) was injected over the surface. Please note that the KD values from both experiments are highly consistent with each other. The SPR analysis is performed by Biaffin GmbH & Co KG, Germany.

Additionally, ACRO also tested their performance in a standard ELISA analysis against Centuximab and 0.2µg/well Centuximab was used to coat the plate(Fig. 5).

Fig. 5 The same two lots of EGFR-biotin (Cat # EGR-H82E7) were used is a standard ELISA assay, Please note that the results were consistent between these two batches.