PROTEOSTAT(R) 蛋白聚集檢測

來源：作者：人氣：-發(fā)表時間：2023-02-28 13:33:00【大中小】

PROTEOSTAT^® 蛋白聚集檢測

某些肽鏈在經(jīng)過錯誤折疊后很容易形成聚集，在生物技術(shù)和制藥領(lǐng)域，這些聚集會嚴重影響蛋白的生產(chǎn)和應(yīng)用。許多以蛋白多肽為基礎(chǔ)研發(fā)的藥物具有極大的應(yīng)用前景，可是蛋白因錯誤折疊而導致構(gòu)象發(fā)生變化形成聚集，使蛋白失活，從而影響藥物療效，造成不可估計的損失。污染物導致的蛋白聚集包括宿主細胞蛋白、非蛋白微粒和蛋白的損傷形式。

基于此 Enzo Life Sciences 推出了新產(chǎn)品 PROTEOSTAT^® Protein Aggregation Assay，內(nèi)含紅色熒光信號染料，可特異性結(jié)合聚集蛋白，進行檢測。具有廣泛的檢測范圍，可有效檢測可見和不可見的蛋白微粒。

◆原理

蛋白聚集嚴重影響以蛋白為基礎(chǔ)研發(fā)的藥物。在藥物制劑中，蛋白聚集在生物活性和免疫原性方面影響效力。

蛋白聚集發(fā)生在生產(chǎn)過程的各個階段包括細胞培養(yǎng)、純化、生產(chǎn)、儲存、運輸?shù)确矫?。制藥工業(yè)希望在生物工藝中找到新的方法，可用于檢測、追蹤、定量分析影響蛋白聚集的因素。

近年來以凍干穩(wěn)定形式存在的蛋白聚集體作為標準品，可以準確地定量檢測蛋白聚集，加上新穎的只需在酶標儀里即可實驗的 PROTEOSTAT^®protein aggregation assay，可對蛋白檢測方法進行優(yōu)化。

在這篇文章中，使用已知濃度的聚集 IgG 標準品作為標準曲線，通過 PROTEOSTAT^® protein aggregation assay/standards 對 IgG 的聚集水平進行檢測，集中分析了制藥工業(yè)中常見的３種聚集形式。

　　● 熱誘導聚集

　　● 機械誘導聚集

　　● 冷凍和反復凍融誘導聚集

◆優(yōu)點·特色

　●　高通量篩選方法用于流式細胞儀平臺監(jiān)測蛋白穩(wěn)定性

　●　在熒光微孔板中即可進行簡單、靈敏、均一地檢測

　●　不需分離目的蛋白或稀釋樣品

　●　與傳統(tǒng)蛋白聚集檢測染料相比更靈敏，信號更明亮

　●　優(yōu)化的緩沖液和輔料用于蛋白制劑研發(fā)

　●　檢測靈敏度可低至亞微摩爾級別，含量在１%-５%的聚集蛋白也能被檢測出

　●　在廣泛的 PH（4-10）和離子強度范圍都可使用，提供至少兩個數(shù)量級的線性動態(tài)范圍

　●　使用 PROTEOSTAT^® 蛋白聚集檢測標準品可對溶液中的聚集蛋白精確定量

　●　可檢測蛋白多肽聚集程度，可用于篩選蛋白聚集激活劑或抑制劑

◆內(nèi)容·使用

試劑盒包含：

PROTEOSTAT®檢測試劑

PROTEOSTAT®陽性對照

PROTEOSTAT®陰性對照

10X PROTEOSTAT®分析緩沖液

在蛋白聚集實驗研究中，將 PROTEOSTAT^® 蛋白聚集檢測試劑盒和 Synergy Mx 多模式微孔板讀數(shù)器聯(lián)用，適用于監(jiān)測由溫度、機械損傷和反復凍融所引起的蛋白聚集。

穩(wěn)定的蛋白顆粒作為標準品可快速建立標準曲線，加快定量分析蛋白聚集響應(yīng)。這個檢測方法能夠可靠地檢測到濃縮抗體制劑中小于 0.2% 的聚集蛋白，線性動態(tài)范圍為２個數(shù)量級。

PROTEOSTAT^® 蛋白聚集檢測試劑盒和 PROTEOSTAT^® 蛋白聚集檢測標準品檢測蛋白聚集特點在于操作簡單，能夠檢測低水平的蛋白聚集情況，只需 30 分鐘，就能夠得到完整的分析結(jié)果。

案例一不同攪拌速率對蛋白聚集的影響	案例二蛋氨酸被氧化后可抑制蛋白聚集
使用本產(chǎn)品檢測后，可知攪拌速度越快，蛋白聚集程度越大。	藍色曲線為對照，沒有攪拌速度在 300 rpm 下，綠色曲線與紅色曲線分別表示蛋氨酸被氧化后的蛋白聚集情況與天然蛋白的聚集情況。使用本產(chǎn)品檢測后，可知蛋氨酸被氧化后可抑制蛋白聚集。

使用說明：

應(yīng)用：流式細胞術(shù)、熒光顯微鏡、熒光檢測和微孔板
該試劑盒專門設(shè)計用于監(jiān)測溶液中蛋白質(zhì)聚集體的形成。

質(zhì)量控制：PROTEOSTAT®蛋白質(zhì)聚集檢測試劑盒用于測定（1）20μM聚集溶菌酶，（2）20μM天然溶菌酶；和（3）5%聚集混合物。Z’因子大于0.5，5%的聚集信號大于無聚集質(zhì)控品的3個標準方差以上。

檢測數(shù)量：
ENZ-51023-KP050：在96孔板中進行50次測試，或流式細胞儀測試16次
ENZ-51023-KP002: 96孔測試2次或流式細胞儀測試70次

儲存：在適當?shù)膬Υ鏃l件下，試劑盒組分在產(chǎn)品標簽注明的日期前穩(wěn)定。將試劑盒儲存在-20°C的無霜冰箱中，或-80°C的條件下長期儲存。

運輸：干冰運輸

監(jiān)管狀態(tài)：RUO - 僅供研究用途

◆參考文獻

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◆使用流式細胞儀結(jié)合PROTEOSTAT^®蛋白聚集分析試劑盒使用例

A PROTEOSTAT^®蛋白聚集分析試劑盒檢測 IgG 聚集標準品 B PROTEOSTAT^®蛋白聚集分析試劑盒檢測硅油滴 C PROTEOSTAT^®蛋白聚集分析試劑盒檢測 IgG 聚集標準品和硅油滴的混合物。

使用流式細胞儀結(jié)合 PROTEOSTAT^®蛋白聚集分析試劑盒檢測 IgG 聚集標準品，可與其他非蛋白微粒如油滴區(qū)別開來，且無須考慮樣品微粒大?。ń?jīng)檢測 >97% 的 IgG 聚集標準品都小于２μm）。

產(chǎn)品編號	產(chǎn)品名稱	產(chǎn)品規(guī)格
ENZ-51023-KP050	PROTEOSTAT^® 蛋白聚集檢測 PROTEOSTAT^® Protein aggregation assay	50 tests
ENZ-51023-KP002	PROTEOSTAT^® 蛋白聚集檢測 PROTEOSTAT^® Protein aggregation assay	2×96 tests

◆相關(guān)產(chǎn)品

產(chǎn)品編號	產(chǎn)品名稱	包裝
ENZ-51039-KP002	PROTEOSTAT^® 蛋白質(zhì)聚集標準品套裝 PROTEOSTAT^® Protein aggregation standards	2x96 wells

◆相關(guān)產(chǎn)品

人IgG多抗

◆產(chǎn)品詳情

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濃度	1mg/ml
來源	兔
免疫原	人免疫球蛋白G
物種反應(yīng)性	人
應(yīng)用	WB
純化細節(jié)	蛋白A親和純化
形式	液體。溶于含有10ppm卡松防腐劑的PBS緩沖液中。
使用/穩(wěn)定性	在4°C下可穩(wěn)定1個月，在-20°C下可穩(wěn)定至少12個月。
處理	避免反復凍融
運輸	冰袋運輸
短期儲存	+4°C
長期儲存	-20°C
監(jiān)管狀態(tài)	RUO - 僅供研究使用

◆產(chǎn)品文獻參考

Th17 micro-milieu regulates NLRP1-dependent caspase-5 activity in skin autoinflammation.; S. Zwicker, et al.; PLoS One 12, e0175153 (2017)

◆產(chǎn)品列表

產(chǎn)品編號	產(chǎn)品名稱	包裝
ENZ-ABS264-1000	人IgG多抗 IgG (human) polyclonal antibody	1 ml

◆相關(guān)產(chǎn)品

PROTEOSTAT® Protein aggregation assay PROTEOSTAT®蛋白聚集分析試劑盒
用于可見及亞可見顆粒蛋白質(zhì)聚集的定量檢測
ENZ-51023-KP002	96 tests×2
ENZ-51023-KP050	50 tests
PROTEOSTAT® Protein aggregation standards PROTEOSTAT®蛋白聚集檢測標準品
市售蛋白質(zhì)聚集測定標準品
ENZ-51039-KP002	2x96 wells
PROTEOSTAT® Thermal shift stability assay kit PROTEOSTAT®熱轉(zhuǎn)移穩(wěn)定性測定試劑盒
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ENZ-51027-K400	400 tests
ENZ-51027-K100	100 tests
PROTEOSTAT® Protein refolding and aggregation sensing kit PROTEOSTAT® 蛋白質(zhì)重折疊和聚集敏感試劑盒
用于確定蛋白質(zhì)重折疊較適宜條件和功能蛋白較大回收率
ENZ-51040-KP002	2x96 wells
Protein A ELISA kit 蛋白A ELISA試劑盒
靈敏的監(jiān)測污染殘留用比色法酶聯(lián)免疫試劑盒（9.01 pg/ml）
ADI-900-057	96 wells
E. coli host cell protein ELISA kit 大腸桿菌宿主細胞蛋白ELISA試劑盒
靈敏的定量檢測大批量大腸桿菌系表達產(chǎn)品的宿主細胞蛋白污染(30 ng/ml)
ENZ-KIT127-0001	96 wells
E. coli host cell protein standards 大腸桿菌宿主細胞蛋白標準品
用于蛋白質(zhì)印跡內(nèi)部對照或ELISA的定量分析的蛋白質(zhì)標準。
ENZ-PRT121-0050	50 μg
E. coli host cell proteins polyclonal antibody 大腸桿菌宿主細胞蛋白多抗
ENZ-ABS262-0100	100 μg
E. coli host cell proteins polyclonal antibody (biotin conjugate) 大腸桿菌宿主細胞蛋白多抗（生物素標記）
ENZ-ABS263-0100	100 μg
CHO host cell protein ELISA kit CHO宿主細胞蛋白ELISA試劑盒
靈敏的定量檢測大批量中國倉鼠（CHO）系表達產(chǎn)品的宿主細胞蛋白污染(10 ng/ml)
ENZ-KIT128-0001	96 wells
CHO host cell protein standards CHO 宿主細胞蛋白標準品
用于蛋白質(zhì)印跡內(nèi)部對照或ELISA的定量分析的蛋白質(zhì)標準。
ENZ-PRT122-0050	50 μg
CHO host cell proteins polyclonal antibody CHO宿主細胞蛋白多抗
ENZ-ABS260-0100	100 μg
CHO host cell proteins polyclonal antibody (biotin conjugate) CHO宿主細胞蛋白生物素標記多抗
ENZ-ABS261-0100	100 μg
IgG (human) polyclonal antibody 人IgG多抗
ENZ-ABS264-1000	1 mL