檢測機構使用流動成像顯微鏡來表征蛋白質聚集體（一）

Contract Lab Using Flow Imaging Microscopy to Characterize Protein Aggregates
檢測機構使用流動成像顯微鏡來表征蛋白質聚集體

Posted by Joyce Brown

Joyce Brown發表

Dan Berdovich knew there were protein aggregates in his customer’s parenteral sample but despite an assortment of high-tech instrumentation at his disposal, he couldn’t be completely confident he was seeing them all or getting a precise count.

Dan Berdovich了解他客戶的腸外樣品中有蛋白質聚集體，但盡管有各種各樣的高科技儀器可供他使用，但他仍然無法完全確信他能看到它們或獲得精確計數。

For an authority on particle analysis who’d earned an award for a peer-reviewed paper on visual inspection standards of injectable drug products, it was disconcerting. Using imaging particle analysis, however, a technology available to industrial markets for less than a decade, Berdovich’s efforts to solve his customers’ problems led to his own business success.

對于顆粒分析的權威機構，他們獲得了關于注射藥物產品目視檢查標準的同行評審論文獎，這令人不安。 然而，使用顆粒成像分析，雖然在工業市場的技術應用不到十年，但是Berdovich努力解決客戶的問題以至他自己的業務獲得成功。

His company, Wheeling, Ill.-based contract laboratory Micro Measurement Laboratories, Inc., does particulate matter testing, particle identification, method validation and visual inspection standards development for pharmaceutical and biotechnology companies world wide, among other services.

他的公司位于伊利諾伊州惠靈的實驗室Micro Measurement Laboratories，Inc。為世界各地的制藥和生物技術公司以及其他公司進行顆粒物測試，顆粒識別，方法驗證和視覺檢測標準開發。

The company offers particular expertise in standards and recommendations set by the U.S. Pharmacopeial Convention (USP), including USP <787>, which covers the testing of sub-visible particulate matter in therapeutic protein injections; USP <788>, which covers testing for and reporting of sub-visible particulate matter in injections and parenteral infusions; and the forthcoming USP <790>, which covers the inspection of visible particulates in injectable drug products; along with the revisions USP <1787>, <1788> and <1790>.

該公司在美國藥典委員會（USP）制定的標準和建議方面提供了特殊的專業知識，包括USP <787>，其中包括治療性蛋白質注射中亞可見顆粒物的測試; USP <788>，其中包括注射和腸胃外輸注中亞可見顆粒物的測試和報告; 以及即將出版的USP <790>，其中包括檢查注射藥品中的可見微粒; 以及USP <1787>，<1788>和<1790>的修訂版。

“I was getting very frustrated that when it came to sub-visible or semi-transparent particles like proteins, we were essentially in the dark,” says Berdovich, “and USP <788> is our bread and butter.”

“我非常沮喪的是，當遇到像蛋白質這樣的亞可見或半透明顆粒時，我們基本上處于黑暗中，”Berdovich說，“USP <788>是我們的面包和黃油。”

Why it hurt
為什么會沮喪

Berdovich’s team used light obscuration instruments to detecting protein aggregates and other particles, as prescribed in USP <788>. The problem, he realized, was that light typically travels through the particles without being scattered. Proteins and protein aggregates, along with other translucent and transparent particles, often defy detection by even the finest light obscuration devices, especially when in the sub-visible range of less than 20 um. Additionally, much larger particles are often severely undersized when measured, if detected at all.

Berdovich的團隊使用光阻法儀檢測蛋白質聚集體和其他顆粒，如USP <788>所述。 他意識到，問題在于光線通常穿過顆粒而不會散射。 蛋白質和蛋白質聚集體以及其他半透明和透明的顆粒，即使是最好的光阻法裝置也經常無法檢測，特別是在小于20微米的亞可見光范圍內。 另外，如果檢測到的話，更大的顆粒通常在測量時測量尺寸會小于實際尺寸。

Characterizing protein aggregates is even more challenging using manual visual inspection-looking at individual vials under controlled lighting conditions and determining whether visible particles are present using the naked eye. This method has limitations similar to those using light obscuration, and because it is inherently subjective, it can yield unreliable results, especially in some biologics and lyophilized drugs.

使用手動目視檢查表征蛋白質聚集體更具挑戰性 - 在受控光照條件下觀察單個樣品瓶，并使用肉眼確定是否存在可見顆粒。 該方法具有類似于使用光阻法的局限性，并且因為其本質上是主觀的，它可以產生不可靠的結果，尤其是在一些生物制劑和凍干藥物中。

At present, 100% visual inspection is required for visible particulates and two methods, light obscuration and membrane microscopy, are accepted by USP to test for the presence of sub-visible particles in parenteral formulations.

目前，可見顆粒需要100％目視檢查，USP接受兩種方法，即光阻法和膜顯微鏡法來測試腸胃外制劑中亞可見顆粒的存在。