Fluorescent-labeled proteins (FTIC and PE-labeled) for biological drug development

Fluorescent-labeled proteins can be robust tools for the detection of antibody screening and CAR expression using flow cytometry. But it is difficult to develop high-quality fluorescent-labeled proteins, and there are very few products available in the market.

ACROBiosystems is a reliable partner to offer antigens for biologic drug discovery and development, developing a unique collection of PE-labeled and FITC-labeled proteins by using the chemical labeling technique developed in-house.

These fluorescent-labeled proteins are exclusively ideal for flow cytometry-based assays, for example, detecting cell-based ligand binding assays and anti-CD19 CAR expression.

Features and benefits

  • Identical binding capacity before and after conjugation
  • Bioactivity verified by flow cytometry and protocol offered
  • High batch-to-batch consistency

Case studies

Evaluation of CAR expression by flow cytometry

FACS analysis of anti-CD19 CAR expression

293 cells were transfected with anti-CD19-scFv and RFP tag. 2e5 of the cells were stained with FITC-Labeled Human CD19 (20-291), Fc Tag (Cat. No. CD9-HF251, 10 μg/mL) and FITC-labeled protein control. Non-transfected 293 cells and FITC-labeled protein control were used as negative control. RFP was used to evaluate CAR (anti-CD19-scFv) expression and FITC was used to evaluate the binding activity of FITC-Labeled Human CD19 (20-291), Fc Tag (Cat. No. CD9-HF251) (QC tested).

293 cells were transfected with anti-CD19-scFv and RFP tag. 2e5 of the cells were stained with FITC-Labeled Human CD19 (20-291), Fc Tag (Cat. No. CD9-HF251, 10 μg/mL) and FITC-labeled protein control. Non-transfected 293 cells and FITC-labeled protein control were used as negative control. RFP was used to evaluate CAR (anti-CD19-scFv) expression and FITC was used to evaluate the binding activity of FITC-Labeled Human CD19 (20-291), Fc Tag (Cat. No. CD9-HF251) (QC tested). Image Credit: ACROBiosystems

Flow cytometry-based ligand binding assay

Flow Cytometry assay shows that PE-Labeled Human PD-1, Fc Tag, His Tag (recommended for neutralizing assay) (Cat. No. PD1-HP2F2) can bind to 293T cells overexpressing human PD-L1. 1 μL stock solution per million cells (QC tested).

Flow Cytometry assay shows that PE-Labeled Human PD-1, Fc Tag, His Tag (recommended for neutralizing assay) (Cat. No. PD1-HP2F2) can bind to 293T cells overexpressing human PD-L1. 1 μL stock solution per million cells (QC tested). Image Credit: ACROBiosystems

Flow cytometry-based neutralizing antibody screening assay

FACS analysis shows that the binding of PE-Labeled Human PD-1, Fc Tag, His Tag (recommended for neutralizing assay) (Cat.No.PD1-HP2F2) to 293T overexpressing PD-L1 was inhibited by increasing concentration of neutralizing antihuman PD-L1 antibody. The IC50 is 0.29 μg/mL (QC tested).

FACS analysis shows that the binding of PE-Labeled Human PD-1, Fc Tag, His Tag (recommended for neutralizing assay) (Cat.No.PD1-HP2F2) to 293T overexpressing PD-L1 was inhibited by increasing concentration of neutralizing antihuman PD-L1 antibody. The IC50 is 0.29 μg/mL (QC tested). Image Credit: ACROBiosystems

High batch-to-batch consistency

Binding activity of three different lots of FITC-labeled Human MSLN, Fc Tag was evaluated in the above FACS analysis against Anti-MSLN CAR-293 cells. The result shows very high batch-to-batch consistency.

Binding activity of three different lots of FITC-labeled Human MSLN, Fc Tag was evaluated in the above FACS analysis against Anti-MSLN CAR-293 cells. The result shows very high batch-to-batch consistency. Image Credit: ACROBiosystems

Binding activity of three different lots of FITC-labeled Human MSLN, Fc Tag was evaluated in the above ELISA analysis against Anti-MSLN Antibody. The result shows very high batch-to-batch consistency.

Binding activity of three different lots of FITC-labeled Human MSLN, Fc Tag was evaluated in the above ELISA analysis against Anti-MSLN Antibody. The result shows very high batch-to-batch consistency. Image Credit: ACROBiosystems

Identical binding capacity before and after conjugation

Binding activity of the Human MSLN, Fc Tag before and after FITC labeling was evaluated in the above ELISA analysis. The result shows that FITC-Labeled and Unconjugated Human MSLN, Fc Tag have almost the same level of binding activity.

Binding activity of the Human MSLN, Fc Tag before and after FITC labeling was evaluated in the above ELISA analysis. The result shows that FITC-Labeled and Unconjugated Human MSLN, Fc Tag have almost the same level of binding activity. Image Credit: ACROBiosystems

Product list

PE labeled protein

Table 1. Source: ACROBiosystems

Molecule Cat. No. Species Product Description Host Structure
BCMA BCA-
HP2H2
Human PE-Labeled Human BCMA / TNFRSF17 Protein, His Tag HEK293
CD19 CD9-
HP2H3
Human PE-Labeled Human CD19 (20-291) Protein, His Tag HEK293
Mesothelin MSN-
HP223
Human PE-Labeled Human Mesothelin (296-580) HEK293
PD-1 PD1-
HP2F2
Human PE-Labeled Human PD-1, Fc Tag, His Tag (recommended for neutralizing assay) HEK293
SIRP alpha SIA-
HP252
Human PE-Labeled Human SIRP alpha, Fc Tag
(recommended for neutralizing assay)
HEK293

 

FITC labeled protein

Table 2. Source: ACROBiosystems

Molecule Cat. No. Host Product Description Structure
BCMA BCA-HF254 HEK293 FITC-Labeled Human BCMA / TNFRSF17 Protein, Fc Tag DMF FiledHot
BCA-HF2H1 HEK293 FITC-Labeled Human BCMA / TNFRSF17 Protein, His Tag DMF Filed
BCA-HF2H3 HEK293 FITC-Labeled Human BCMA / TNFRSF17 Protein, His Tag Star Staining
CD19 CD9-HF251 HEK293 FITC-Labeled Human CD19 (20-291) Protein,
Fc Tag DMF FiledHot
CD9-HF2H2 HEK293 FITC-Labeled Human CD19 (20-291) Protein, His Tag DMF FiledHot
CD9-HF2H3 HEK293 FITC-Labeled Human CD19 (20-291) Protein, His Tag Star Staining

 

Click here to learn about many more FITC labeled proteins.