バイオサイトジェンでは細胞と生化学分析サービスを全面的に提供しております。In vitro測定実験には初代細胞と継代細胞測定に分けています。初代細胞の測定にはアゴニスト誘導のT細胞活性化、混合リンパ球反応（MLR）試験、抗原の回収試験、T細胞およびNK細胞毒性試験、DC細胞活性化試験、マクロファージ貪食試験、抗体依存性細胞毒性（ADCC）試験、相補体依存性細胞毒性（CDC）試験、および抗体結合実験が含まれます。通常サイトカイン産生試験やフローサイトメトリー法（FACS）で細胞マーカーを測定します。継代細胞の測定には、細胞表面マーカーの発現、抗体の結合などが含まれ、細胞増殖や細胞死の統計、蛍光や生物発光あるいは光学系を用いたレポーターシステム遺伝子アッセイがあります。バイオサイトジェンはまた表面プラズモン共鳴（SPR）を用いて、生化学的な結合測定や親和性の測定も行われています。
弊社のサービス
薬物の細胞結合測定
薬物とヒト化マウス脾臓細胞の結合測定
薬物阻害試験
in vitro刺激試験
MLRのin vitro刺激試験
in vitro細胞毒性試験（ADCC,CDC)
溶血/凝血の試験
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細胞表現抗原結合試験：MC38-ｈPD-L1細胞表面でのhPD-L1の測定
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MC38 is a mouse colon cancer cell line. MC38-hPD-L1 is an engineered MC38 cell line expressing human PD-L1 in place of mouse PD-L1. Analysis of PD-L1 expression with anti-mouse and anti-human PD-L1 antibodies by flow cytometry demonstrates that human PD-L1 indeed is expressed only in MC38-hPD-L1, whereas mouse PD-L1 is only expressed in the wild type MC38
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事例2：抗体依存性細胞毒性（ADCC）測定
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Ratio=(CFSElow cell /CFSEhigh cell)
Specific Cytotoxicity=[1-(No-drug control Ratio/Experimental Ratio)]*100%
Fig 1. ADCC with anti-human CTLA4 antibody
In this assay, human PBMCs were used as effector cells. Jurkat- hCTLA4 cells were labeled with high concentration of CFSE as the target cells, and Jurkat WT cells labeled with low concentration of CFSE as internal controls. Effector cells, target and internal control cells were incubated for 18 hours in the presence of varied concentrations of anti-human CTLA-4. Specific Cytotoxicity=[1-(No-drug control Ratio/Experimental Ratio)]*100%; Ratio=(CFSElow cells /CFSEhigh cells)
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事例3：抗体依存性細胞毒性（CDC）測定
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リツキシマブのモノクローナル抗体介在のCDC実験です。補体の含む培養基で培養するRaji細胞に異なる濃度のリツキシマブ（モノクローナル抗体）を入れて2時間培養し、培養基中に細胞から放出されたLDHを細胞毒性の指標として測定します。リツキシマブ（モノクローナル抗体）はRaji細胞に対して強いCDC活性化を表します。
細胞毒性%＝（実験ウェルLDH測定値-自発的放出のウェルLDH測定値）/（細胞の最大LDH放出測定値）*100%
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事例4：T細胞活性化試験
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Human PBMCs (0.1 million) were added into wells of a round bottom 96-well culture plate with SEB (10 ng/mL) and varied amount of Ipilimumab or isotype control, and incubated for 48 hours. Release of IL-2 was determined by ELISA. The results show that ipilimumab dose-dependently potentiated SEB stimulated IL-2 production by PBMCs.
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事例5：混合リンパ球反応(MLR)測定
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Human T cells and allogeneic DCs were mixed in the ratio of 5:1 in wells of a round bottom 96-well culture plate in the presence of varied concentration of nivolumab or isotype control antibody and incubated for 72 hours. Release of IFN-γ was determined by ELISA. The results show that nivolumab dose-dependently potentiated IFN-γ production in mixed lymphocyte reaction (MLR).
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事例6：赤血球沈降試験
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Two percent (2%) red blood cells (RBC) was incubated with anti-human CD47 antibody of varied concentrations from 0 ng/mL to 10 ng/mL. Erythrocyte sediment was detected after 0.5 hours. RBC would normally precipitate to the bottom without toxic or pathological interference. As shown in the panel, toxic anti-CD47 antibody (top) prevented RBC sediment at lower threshold concentration (indicated by the red bars) than the less toxic anti-CD47 antibody (bottom).
