xCELLigence® Immunotherapy Kits Liquid Tumor Killing Assays

Continuously monitor liquid cancer killing with immunotherapy kits compatible with xCELLigence Real Time Cell Analyzers.

Monitor the Efficacy of Liquid Cancer Immunotherapies inside your incubator

Use xCELLigence Immunotherapy Kits with your real-time cell analysis (RTCA®) system for a non-invasive solution to a broad range of liquid cancer immunotherapies and suspension tumor cell killing applications.

  • Label-Free: Allowing for more physiological assay conditions; labeling or secondary assays aren’t required
  • Real-Time: Quantitative monitoring of both fast (hours) and slow (days) killing kinetics
  • Sensitive: Capable of evaluating low effector cell to target cell ratios that are physiologically relevant
  • Simple Workflow: Requires only the addition of effector cells to target cells (in the presence or absence of antibodies); homogeneous assay without additional sample handling
  • Automatic Data Plotting: RTCA software Pro enables facile data display and objective analysis, precluding the subjective data vetting that is common to imaging-based assays


The high specificity and potent cytotoxicity of immune system effector cells make them promising agents for extirpating liquid cancers.  Though the list of efficacious cell-mediated immunotherapies is growing, realizing the full therapeutic potential of this field will require: (1) continued elucidation of the mechanisms underlying cancer cell recognition and immune cell-mediated killing, and (2) an ability to screen immunotherapy constructs/conditions using patient-derived effector and/or target cells to identify optimal treatment regimens.  Fundamental to both of the above is the ability to quantitatively monitor the potency of immune cell-mediated killing of target liquid cancer cells under controlled conditions in vitro.  Traditional cell killing assays suffer from drawbacks that prevent them from meeting this need efficiently.

ACEA’s xCELLigence Real-Time Cell Analysis (RTCA) instruments utilize gold microelectrodes embedded in the bottom of microtiter wells to non-invasively monitor the status of adherent cells. Parameters analyzed include cell number, cell size/shape, and cell-substrate attachment strength. Though they are not naturally adherent, liquid cancer cells can be immobilized on the plate bottom by pre-coating the wells with a tethering antibody. Upon exposure to diverse immunotherapies (NK cell, T cells, CAR-T, oncolytic virus, checkpoint inhibitors, etc.) the target cell response is automatically recorded by the xCELLigence instrument as shown in detail below. The major distinguishing features of this technology include enhanced sensitivity, the preclusion of labels, simple workflow and, importantly, continuous kinetic measurement of target liquid cancer cell health/behavior.Immunotherapy Kits - Liquid Tumor Workflow

Step 1: The target liquid cancer cells are first seeded in the wells of an electronic microtiter plate (E-Plate®) that have been pre-coated with a tethering antibody.  Interaction of the seeded cells with the gold microelectrodes impedes the flow of electric current between electrodes.  This impedance value, plotted as a unitless parameter called “Cell Index”, increases as cells proliferate and then plateaus as cells approach 100% confluence.

Step 2: When added subsequently, non-adherent effector cells (i.e. immune cells) in suspension do not cause impedance changes in and of themselves (due to lack of adherence to the gold microelectrodes).

Step 3: If effector cells and/or antibodies induce the destruction of the target cells, this cytolytic activity can be sensitively and precisely detected.  The continuous acquisition of impedance data for each well of an E-Plate enables the generation of real-time killing curves for multiple conditions simultaneously.

xCELLigence Immunotherapy Kits Brochure Download PDF

ACEA has developed immunotherapy kits with five different tethering specificities to enable the evaluation of immunotherapies targeting various types of liquid cancers:
ProductTethering SpecificityValidated Effector Cells†Validated Target Cells†
Liquid Tumor Killing Assay (anti-CD40)
anti-CD40NK-92
CAR-T
primary CD8+ T cells
Daudi
Raji
Ramos
MEC2
Liquid Tumor Killing Assay (anti-CD19)
anti-CD19NK-92
primary CD8+ T cells
Raji
Liquid Tumor Killing Assay (anti-CD29)anti-CD29NK-92*K562
HEL 92.1.7
Liquid Tumor Killing Assay (anti-CD9)anti-CD9NK-92NALM6
RS4;11
RPMI 8226
Liquid Tumor Killing Assay (anti-CD71)anti-CD71NK-92K562
THP-1

The cell types listed here have already been successfully utilized.  However, the kit is amenable to use with many other cell types.
*Because NK cells express low levels of CD29, they can be tethered by the anti-CD29 tethering reagent that is used in this assay.  Although the impedance signal resulting from NK cells can be subtracted using the xIMT software, the kit is most effective when using alternative effector cell types such as T cells, CAR-T, etc.).

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xCELLigence Instruments for Immunotherapy Assays:
xCELLigence RTCA DPxCELLigence RTCA SPxCELLigence RTCA MPxCELLigence RTCA HT
RTCA DPRTCA SPRTCA MPRTCA HT
3×16 wells1×96 wells6×96 wellsUp to 4×384 wells


Liquid Tumor Killing Assay (anti-CD40)

Complete Kit (Cat# 8100004)Tethering Kit (Cat# 8100005)Sample Kit (Cat# 8100006)
E-Plate View 96 (Qty=6)Tethering Reagent (anti-CD40) (250 µL)E-Plate View 96 (Qty=2)
Tethering Reagent (anti-CD40) (250 µL)10X Tethering Buffer (10 ml)Tethering Reagent (anti-CD40) (90 µL)
10X Tethering Buffer (10 ml)Cytolysis Reagent (1.5 ml)10X Tethering Buffer (10 ml)
Cytolysis Reagent (1.5 ml)Cytolysis Reagent (1.5 ml)
RTCA Software ProTrial RTCA Software Pro for 1-month usage

Liquid Tumor Killing Assay (anti-CD29)

Complete Kit (Cat# 8100007)Tethering Kit (Cat# 8100008)Sample Kit (Cat# 8100008)
E-Plate View 96 (Qty=6)Tethering Reagent (anti-CD29) (125 µL)E-Plate View 96 (Qty=2)
Tethering Reagent (anti-CD29) (125 µL)10X Tethering Buffer (10 ml)Tethering Reagent (anti-CD29) (45 µL)
10X Tethering Buffer (10 ml)Cytolysis Reagent (1.5 ml)10X Tethering Buffer (10 ml)
Cytolysis Reagent (1.5 ml)Cytolysis Reagent (1.5 ml)
RTCA Software ProTrial RTCA Software Pro for 1-month usage

Liquid Tumor Killing Assay (anti-CD19)

Complete Kit (Cat# 8100010)Tethering Kit (Cat# 8100011)Sample Kit (Cat# 8100012)
E-Plate View 96 (Qty=6)Tethering Reagent (anti-CD19) (250 µL)E-Plate View 96 (Qty=2)
Tethering Reagent (anti-CD19) (250 µL)10X Tethering Buffer (10 ml)Tethering Reagent (anti-CD19) (90 µL)
10X Tethering Buffer (10 ml)Cytolysis Reagent (1.5 ml)10X Tethering Buffer (10 ml)
Cytolysis Reagent (1.5 ml)Cytolysis Reagent (1.5 ml)
RTCA Software ProTrial RTCA Software Pro for 1-month usage

Liquid Tumor Killing Assay (anti-CD9)

Complete Kit (Cat# 8100013)Tethering Kit (Cat# 8100014)Sample Kit (Cat# 8100015)
E-Plate View 96 (Qty=6)Tethering Reagent (anti-CD9) (250 µL)E-Plate View 96 (Qty=2)
Tethering Reagent (anti-CD9) (250 µL)10X Tethering Buffer (10 ml)Tethering Reagent (anti-CD9) (90 µL)
10X Tethering Buffer (10 ml)Cytolysis Reagent (1.5 ml)10X Tethering Buffer (10 ml)
Cytolysis Reagent (1.5 ml)Cytolysis Reagent (1.5 ml)
RTCA Software ProTrial RTCA Software Pro for 1-month usage

Liquid Tumor Killing Assay (anti-CD71)

Complete Kit (Cat# 8100016)Tethering Kit (Cat# 8100017)Sample Kit (Cat# 8100018)
E-Plate View 96 (Qty=6)Tethering Reagent (anti-CD71) (250 µL)E-Plate View 96 (Qty=2)
Tethering Reagent (anti-CD71) (250 µL)10X Tethering Buffer (10 ml)Tethering Reagent (anti-CD71) (90 µL)
10X Tethering Buffer (10 ml)Cytolysis Reagent (1.5 ml)10X Tethering Buffer (10 ml)
Cytolysis Reagent (1.5 ml)Cytolysis Reagent (1.5 ml)
RTCA Software ProTrial RTCA Software Pro for 1-month usage