PROTEOSTAT® 蛋白聚集檢測某些肽鏈在經過錯誤折疊后很容易形成聚集，在生物技術和制藥領域，這些聚集會嚴重影響蛋白的生產和應用。許多以蛋白多肽為基礎研發的藥物具有極大的應用前景，可是蛋白因錯誤折疊而導致構象發生變化形成聚集，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 推出了新產品 PROTEOSTAT® Protein Aggregation Assay，內含紅色熒光信號染料，可特異性結合聚集蛋白，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研發的藥物。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。
蛋白聚集發生在生產過程的各個階段包括細胞培養、純化、生產、儲存、運輸等方面。制藥工業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
◆優點·特色
● 高通量篩選方法用于流式細胞儀平臺監測蛋白穩定性
● 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
● 與傳統蛋白聚集檢測染料相比更靈敏，信號更明亮
● 優化的緩沖液和輔料用于蛋白制劑研發
● 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，提供至少兩個數量級的線性動態范圍
● 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
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◆內容·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 多模式微孔板讀數器聯用，適用于監測由溫度、機械損傷和反復凍融所引起的蛋白聚集。
穩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。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，線性動態范圍為２個數量級。
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果。
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案例一
 不同攪拌速率對蛋白聚集的影響
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使用本產品檢測后，可知攪拌速度越快，蛋白聚集程度越大。
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案例二
 蛋氨酸被氧化后可抑制蛋白聚集
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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。使用本產品檢測后，可知蛋氨酸被氧化后可抑制蛋白聚集。
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使用說明：
應用：流式細胞術、熒光顯微鏡、熒光檢測和微孔板
該試劑盒專門設計用于監測溶液中蛋白質聚集體的形成。
該試劑盒專門設計用于監測溶液中蛋白質聚集體的形成。
質量控制：PROTEOSTAT®蛋白質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控品的3個標準方差以上。
檢測數量：
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
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：在適當的儲存條件下，試劑盒組分在產品標簽注明的日期前穩定。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
監管狀態：RUO - 僅供研究用途
◆參考文獻
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◆使用流式細胞儀結合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
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B PROTEOSTAT® 蛋白聚集分析試劑盒檢測硅油滴
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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。
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使用流式細胞儀結合 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別開來，且無須考慮樣品微粒大小（經檢測 >97% 的 IgG 聚集標準品都小于２μm）。
| 產品編號 | 產品名稱 | 產品規格 |
| ENZ-51023-KP050 | PROTEOSTAT® 蛋白聚集檢測 PROTEOSTAT® Protein aggregation assay |
50 tests |
| ENZ-51023-KP002 | 2×96 tests |
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