항체치료제 ADCC Assay 서비스

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항체치료제 ADCC Assay 서비스

Maker : EUROFINS DISCOVERY

항체 면역항암제의 효능을 평가하기 위한 ADCC분석서비스를 제공합니다.

프린트 제품 문의

특징

ADCC(Antibody-dependant-cell mediated cytotoxicity, 항체 의존 세포 매개 세포 독성)는 암이나 바이러스가 감염된 타겟 세포를 용해하기 위한 면역세포의 활성화를 유도하는 세포 매개 면역 방어 메커니즘입니다.
항체가 암세포에 결합하여 NK 세포를 활성화시키고, 이로 인해 암세포가 사멸됩니다.
Anti-HER2 ADCC assay (#300230-1)의 경우, BT-474 표적 세포와 PBMC effector cells을 이용하여 타겟 세포에 대한 cytotoxicity 를 형광으로 측정합니다. 공여자 의존성 변이의 영향을 줄이기 위해 3명의 donor로부터 얻은 PBMC를 이용하며, 검증된 Anti-HER2 Drug인 Trastuzumab를 control로 사용합니다.
다양한 표적 및 effector cell을 이용한 맞춤형 ADCC 분석을 개발해 드립니다.

ADCC 원리

데이터

Figure 1. Flow cytometry assessment of HER2 surface expression on three breast cancer cell lines.

MDA-MB-231, MCF-7 and BT-474 cells were thawed and stained with either anti-Her2-PE antibody or isotype control for 30 min followed by washing with 0.2% BSA in PBS. The HER2 surface expression was evaluated from 10,000 cells/sample by flow cytometry. BT-474 cells have a high expression level for HER2 which makes them an ideal target cell.

Figure 2. Cellular cytotoxicity assays enable efficacy comparisons of trastuzumab and a biosimilar.

Specific cytotoxicity is determined from percent cytotoxicity measured from an antibody of interest and an isotype matched control. Percent Cytotoxicity is calculated as: Cytotoxicity % = 100 x (1-(A-C)/ (D-C)) where A = Fluorescence intensity of test compound or reference treated group, C = Fluorescence intensity of medium only, D = Fluorescence intensity of target cell (BT-474) only.

Figure 3. Specific anti-HER2 antibody-dependent cellular cytotoxicity in BT-474 cells.

Fluorescence intensity decreased by ≥ 20% relative to 1 μg/ml hIgG1 isotype control indicates specific ADCC activity. Percent ADCC increases, up to a point, with increasing concentrations of trastuzumab, but not IgG. Antibodies were preincubated with Calcein AM-stained target cells (1 x 104 cells/well) for 30 min, followed by addition of PBMC effector cells. Live cell images were acquired using the ImageXpress Micro Confocal Image System; target cell cytotoxicity was quantified by fluorescence intensity. ADCC is calculated as: 100 x (1-(A-C)/ (B-C)) where A = Fluorescence intensity of test compound or reference treated group, B = Fluorescence intensity of 1 μg/ml human IgG1 treated group, C = Fluorescence intensity of medium only.