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抗獨特型抗體

背景
抗獨特型抗體(anti-idiotypic antibody)是能夠識別另一抗體可變區,并產生特異性結合的抗體。抗獨特型抗體在藥物開發中的應用非常廣泛,可以作為免疫原性(immunogenicity)分析的重要參照,同時也可以特異性檢測體內抗體藥水平,是藥代動力學(pharmacokinetics)研究的重要試劑。
為了支持上述研究,ACROBiosystems公司開發了一系列高親和力、高特異性的抗獨特型抗體,應用于免疫原性分析和藥代動力學研究。對于每一個抗獨特型抗體,我們都會根據不同的應用場景開發相應的實驗方案,希望最大程度上加速藥物研發的進程。目前產品已覆蓋adalimumab, rituximab, cetuximab, trastuzumab, bevacizumab等多個已上市的熱門抗體藥。
產品列表
Cat.No. Antigen Neutralizing Activity Affinity KD, nM Application
TRB-Y1 Trastuzumab F(ab')2 Neutralizing Antibody 0.296 PK bridging ELISA with TRB-Y5b
Neutralizing assay
Indirect ELISA
TRB-Y7 Trastuzumab F(ab')2 Neutralizing Antibody 0.452 ADA assay
Neutralizing assay
Indirect ELISA
TRB-Y5b Trastuzumab F(ab')2 Neutralizing Antibody / PK bridging ELISA with TRB-Y1
ADB-Y19 Adalimumab F(ab')2 Neutralizing Antibody 0.0013 ADA assay
Neutralizing assay
Indirect ELISA
ADB-Y23 Adalimumab F(ab')2 Non-Neutralizing Antibody / PK bridging ELISA with ADB-BY17
Indirect ELISA
ADB-BY17 Adalimumab F(ab')2 Neutralizing Antibody / PK bridging ELISA with ADB-Y23
CEB-Y27 Cetuximab F(ab')2 Neutralizing Antibody 0.007 ADA assay
Neutralizing assay
Indirect ELISA
CEB-Y28 Cetuximab F(ab')2 Neutralizing Antibody 0.0015 ADA assay
Neutralizing assay
Indirect ELISA
CEB-Y29 Cetuximab F(ab')2 Neutralizing Antibody / PK bridging ELISA with CEB-BY31
Neutralizing assay
Indirect ELISA
CEB-Y31 Cetuximab F(ab')2 Non-Neutralizing Antibody 0.421 ADA assay
Indirect ELISA
CEB-BY31 Cetuximab F(ab')2 Non-Neutralizing Antibody / PK bridging ELISA with CEB-Y29
RIB-Y35 Rituximab F(ab')2 Neutralizing Antibody 0.03 ADA assay
Neutralizing assay
Indirect ELISA
RIB-Y36 Rituximab F(ab')2 Neutralizing Antibody 0.01 ADA assay
Neutralizing assay
Indirect ELISA
RIB-Y37 Rituximab F(ab')2 Neutralizing Antibody / PK bridging ELISA with RIB-BY35
Neutralizing assay
Indirect ELISA
BEB-Y12 Bevacizumab F(ab')2 Neutralizing Antibody 0.0828 Neutralizing assay
Indirect ELISA
BEB-Y9 Bevacizumab F(ab')2 Neutralizing Antibody 1.92 ADA assay
Indirect ELISA
BEB-Y10 Bevacizumab F(ab')2 Neutralizing Antibody / PK bridging ELISA with BEB-BY13
Neutralizing assay
Indirect ELISA
BEB-BY13 Bevacizumab F(ab')2 Neutralizing Antibody / PK bridging ELISA with BEB-Y10
應用
幾乎所有的生物制品都會產生一定的抗藥抗體(anti-drug antibody, ADA),ADA的產生可能會降低藥物療效或導致嚴重的不良反應。臨床前研究表明,ADA能對藥物暴露、藥物毒性作用、藥物代謝動力學、藥物效應動力學等造成影響。為了評估生物制品分子的免疫原性,以及將實驗結果與臨床事件聯系起來,在臨床前研究和臨床研究中,有必要開發可靠的能夠有效評估ADA反應的實驗方法。
ADA的檢測方法通常是非定量實驗,因為沒有可用的標準化的ADA作為校正標準品。陽性對照一般都是內部開發的單克隆抗體或多克隆抗體,非常耗時。為了解決這一問題,ACROBiosystems開發了一系列ADA作為標準品,用于對應抗體及其生物類似藥的ADA檢測。
應用案例1- Anti-Rituximab Antibodies (RIB-Y36

Anti-Rituximab Antibodies bridging ELISA for Anti-Drug Antibody (ADA) assay development. Immobilized rituximab at 1 μg/ml, added increasing concentrations of Anti-Rituximab Antibodies (Cat. No. RIB-Y36, 10% human serum) and then added biotinylated rituximab at 2 μg/ml. Detection was performed using HRP-conjugated streptavidin with a sensitivity of 9.7 ng/mL. 国产视频app

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Anti-Rituximab Antibodies bridging MSD for Anti-Drug Antibody (ADA) assay development. Added the mix solution (biotinylated Rituximab at 5 μg/mL, SULFO-Rituximab at 5Nμg/mL and increasing concentrations of Anti-Rituximab Antibodies (Cat. No. RIB-Y36, 100% human serum). Detection was performed using MSD Assay with a sensitivity of 0.97 ng/mL. s

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應用案例2- Anti-Adalimumab Antibodies (ADB-Y19

Anti-Adalimumab Antibodies bridging ELISA for Anti-Drug Antibody (ADA) assay development. Immobilized adalimumab at 1 μg/ml, add increasing concentrations of Anti-Adalimumab Antibodies (Cat. No. ADB-Y19, 10% human serum) and then add biotinylated adalimumab at 5 μg/ml. Detection was performed using HRP-conjugated streptavidin with a sensitivity of 0.6 ng/mL.

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Measured by its neutralizing ability in a functional ELISA. Immobilized adalimumab at 0.5 μg/mL (100 μL/well) can bind pre-mixed Anti-Adalimumab Antibodies (Cat. No. ADB-Y19) and Biotinylated Human TNF-alpha (Cat. No. TNA-H82E3) with an inhibition rate of 100%.

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PK assay -特異性檢測體內抗體藥水平
ELISA是抗體藥血藥濃度檢測中最常用的技術,而indirect ELISA是其中最為簡單、常見的方法。該方法具有操作簡單、靈敏度高等優點。但是,由于二抗通常選擇Anti-Human IgG,導致非特異背景較高,必須對基質稀釋1000倍以上才能避免基質干擾。為了解決這一問題,我們用抗獨特型抗體來檢測基質中的抗體藥水平。具體來講,采用bridging ELISA形式,用抗獨特型抗體對來檢測抗體藥水平,能顯著降低非特異背景。

Comparison between anti-idiotypic capture ELISA and anti-idiotypic bridging ELISA for rituximab detection in patient samples. Left: anti-idiotypic capture ELISA; Right: anti-idiotypic bridging ELISA. 国产视频app

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Detection of rituximab by bridging ELISA in serum. Immobilized Anti-Rituximab Antibodies (Cat. No. RIB-Y37) at 2 μg/ml, added increasing concentrations of Rituximab (10% human serum) and then added biotinylated Anti-Rituximab Antibodies (Cat. No. RIB-BY35) at 1 μg/ml. Detection was performed using HRP-conjugated streptavidin with a sensitivity of 1 ng/ml.

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產品特點
高親和力
親和力代表了抗原和抗體間的結合強度,用KD值表示,KD值越小,抗體的親和力越強。我們用SPR方法檢測了所有抗獨特型抗體的親和力,了解這一指標,有助于在方法學建立過程中預測方法的靈敏度。

Anti-Adalimumab Antibodies (mouse IgG1, Cat. No. ADB-Y19) captured on CM5 chip via anti-mouse antibodies surface, can bind human adalimumab with an affinity constant of 1.36 pM.

高特異性
對于每一個產品,我們都進行了特異性檢測,包括對應的抗體藥、抗體藥靶點、不同IgG亞型和同亞型其他抗體藥,只有結合對應的抗體藥,而不結合其他蛋白或抗體的產品才會被放行。

Demonstration of the specificity of Anti-Cetuximab Antibodies (Cat. No. CEB-Y28) to the cetuximab.

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高穩定性
為確保產品穩定性,ACROBiosystems會做加速實驗和凍融實驗來驗證產品穩定性,只有經過測試證實質量和活性沒有任何影響的產品才會被放行。

Reconstituted Anti-Trastuzumab Antibodies were diluted to 0.4 mg/ml, aliquoted and placed at 37°C. Aliquots were removed from 37°C at every time point and placed at 4°C along with the control. No significant loss of activity was observed.

Anti-Trastuzumab Antibodies were subjected to the indicated number of freeze-thaw cycles (FT). No significant loss of activity was observed.

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