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Key Features

  • Over 200 tumor cell lines, over 200 target-overexpressing stable cell lines, over 10 reporter gene cell lines
  • Over 40 bispecific antibody bioassay projects delivered

Cell-engager

Cell-engager bsAb can active and re-direct effector cell (cytotoxicity T cell, NK cell, etc.) to tumor associated antigen (TAA) cells, and result in the eradication of the malignant cell.

Based on MoA of cell-engager, ProBio offers target-overexpressing cell lines and diverse immune cells to evaluate the biological activity of bsAb in vitro.

Types of bsAbs Target Cell Targets on Target Cell Effector Cell Targets on Effector Cell In Vitro Assay
T cell engager 1. Target overexpressing engineered cell line
2. Tumor cell line
TAA/TSA* 1. 1. Primary cell(T cell /NK cell/macrophage/DC etc.)
2. Reporter cell line
CD3 1. Primary immune cell assay (TDCC etc.)
2. Cytokines release assay
3. Reporter cell line assay
NK cell engager NK cell activating receptors (CD16A, NKG2D, NKp46, NKp30 etc.) 1. Primary immune cell assay (tumor cell killing with primary NK etc.)
2. Reporter cell line assay
Anti-TAA x co-stimulatory immune checkpoint Co-stimulatory immune checkpoint (CD40, 4-1BB, CD28 etc.) 1. Reporter cell line assay
2. Primary immune cell assay (immune cell activation, tumor cell killing, mixed lymphocyte reaction (MLR) etc.)
Anti-TAA x TAA/co-inhibitory immune checkpoint NK cell activating receptor CD16A, macrophage activating receptor CD32A 1. Reporter cell line assay
2. Primary immune cell assay (ADCP etc.)

* Tumor-associated antigens (TAA), which have elevated levels on tumor cells, but are also expressed at lower levels on healthy cells. Tumor-specific antigens (TSA), found on cancer cells only, not on healthy cells.

  • 1. TDCC assay

    case

    Anti-CD3×TAA bsAb induced TAA-expressing cell lysis in a dose-dependent manner.

  • 2. Reporter cell line assay

    case

    Anti-CD3×TAA bsAb activated T cell in a dose-dependent manner.

 

Case 2: Characterizations of Anti-TAA x co-stimulatory immune checkpoint bsAb

Anti-4-1BB×TAA bsAb reporter cell line assay

  • case
  • case

Anti-4-1BB×TAA bsAb significantly activated 4-1BB signaling in reporter cells upon co-culture with TAA-overexpressing CHO-K1 cells in a dose-dependent manner (left). However, anti-4-1BB×TAA bsAb showed weak activation of 4-1BB signaling when co-incubated with CHO-K1 cells (right).

 

Dual-target blockade

As a potent immunotherapy for autoimmune diseases and cancer, blockading multiple pathological factors and pathways by bsAb is a promising approach in improved therapeutic efficacy.

Types of bsAbs Targets In Vitro Assay
Blocking angiogenesis and/or tumorigenesis VEGF & ANG2, Her2 & Her2, EGFR & c-Met, VEGF & DLL4 etc. 1. Cell signaling pathway assay
2. Reporter cell line assay
3. Enzyme activity assay
4. Cytokines/chemokines release assay
5. Antibody internalization assay
6. Cell proliferation assay
7. Primary immune cell assay (ADCC, MLR etc.)
Modulating tumor microenvironment (TME) TGFβ & PDL1, TGFβ & CD73, TGFβ & CD39 etc.
Targeting inflammatory/autoimmune diseases TNFα & IL17A, IL4 & IL13, BAFF & IL17A etc.
Blocking angiogenesis and/or tumorigenesis x Anti-immune checkpoint VEGF & PD1, EGFR & PD1, Her2 & PD1 etc.

Case 1: Blocking angiogenesis and/or tumorigenesis bsAb in vitro assay

VEGF & ANG2 cell signaling pathway assay

  • case
  • Compared with anti-ANG2 mAb or anti-VEGF mAb treatment alone, combination of anti-ANG2 mAb and anti-VEGF mAb showed synergetic reduction in p-AKT level induced by ANG2 and VEGF. This bioassay is suited to evaluate characterization of anti-ANG2×VEGF bsAb.

Case 2: Targeting inflammatory/autoimmune diseases bsAb in vitro assay

TNFα & IL17A chemokines release assay

  • case
  • Compared with anti-TNFα mAb or anti-IL17A mAb treatment alone, combination of anti-TNFα mAb and anti-IL17A mAb could more effectively inhibite TNF-α and IL-17a mediated CXCL1 production. This bioassay is suited to evaluate characterization of anti-TNFα×IL17A bsAb.

Dual immunomodulators

Because combination of immune checkpoint inhibitors have shown significant success in promoting immune responses against cancer and can result in tumor regression in many patients, bsAb targeting two immune checkpoints to activate immune cell has quickly become a research hotspot.

Based on MOA of dual immunomodulators, ProBio develops dual target reporter gene systems and primary immune cell assays to verify the synergistically bioactivities of bsAb compared to the treatment alone or combination of monovalent monoclonal antibody.

Types of bsAbs Targets In Vitro Assay
Anti-co-inhibitory immune checkpoint × co-inhibitory immune checkpoint PD1 & TIGIT, PD1 & CTLA4 ect. 1. Reporter cell line assay
2. Primary immune cell assay (immune cell activation, MLR etc.)
Anti-co-inhibitory immune checkpoint × co-stimulatory immune checkpoint PD1 & 4-1BB, PD1 & OX40 ect.
Anti-co-stimulatory immune checkpoint × co-stimulatory immune checkpoint 4-1BB & CD40, 4-1BB & OX40 ect.
  • Case 1: Anti-co-inhibitory immune checkpoint × co-inhibitory immune checkpoint bsAb in vitro assay

    PD-1+CTLA4 combination bioassay

    case

    Compared with anti-PD-1 mAb or anti-CTLA4 mAb treatment alone, combination of anti-PD-1 mAb and anti-CTLA4 mAb could more effectively activated PD-1&CTLA4 reporter cell line. This bioassay is suited to evaluate characterization of anti-PD-1×CTLA4 bsAb.

  • Case 2: Anti-co-inhibitory immune checkpoint × co-stimulatory immune checkpoint bsAb in vitro assay

    PD-1+4-1BB combination bioassay

    case

    Compared with anti-PD-L1 mAb or anti-4-1BB mAb treatment alone, combination of anti-PD-L1 mAb and anti-4-1BB mAb could more effectively activated 4-1BB&PD-1/PD-L1 reporter cell line. This bioassay is suited to evaluate characterization of anti-4-1BB×PD-1/PD-L1 bsAb.

Cell surface protein bridging

Through clustering targeting molecules to form antigen complexes and aggregations, protein bridging bsAbs can mimicking effects of natural agonist to activates signaling cascade (i.e., HEMLIBRA® restore the blood clotting cascade).

Based on MoA of cell surface protein bridging, ProBio offers customized approaches to evaluate your biological candidates.

Types of bsAbs Targets In Vitro Assay
Apoptotic signaling pathway DR5 & TAA etc. Cell activity assay
Coagulation pathway Factors IXa & Factors X APTT (activated partial thromboplastin time) assay

Case :  Characterization of apoptosis activating bsAb in vitro assay

case

Compared with anti-DR5 mAb treatment, anti-TAA×DR5 bsAb could kill tumor cells at low concentration (tumor cell growth inhibition rate at 0.002nM close to 80% ).

Bispecific Antibody Discovery Services

ProBio provides one-stop bsAb discovery from target validation to preclinical candidates(PCC) , including bsAb format design for different MOA and target , bsAb construction and expression, functional validation, in vivo assay and developability.

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