BioVector? CLM24 糖工程大腸桿菌底盤菌株 / BioVector? CLM24 Glycoengineered E. coli Chassis Strain

通用定義 / General Definition:

BioVector? CLM24 是一種經(jīng)過基因工程高度優(yōu)化的大腸桿菌 (Escherichia coli) 底盤菌株，專門用于蛋白質(zhì)糖基化 (Protein Glycosylation) 的研究與生產(chǎn)。該菌株源自經(jīng)典的 $E. coli$ W3110，其核心改造是徹底敲除了 $waaL$ 基因（$O$-抗原連接酶基因）。通過阻斷內(nèi)源性 $O$-抗原向脂多糖 (LPS) 的轉(zhuǎn)移，CLM24 能夠?qū)⒅|(zhì)載體上的聚糖底物高效地“讓渡”給異源表達(dá)的寡糖轉(zhuǎn)移酶（如來自空腸彎曲桿菌的 PglB），從而實現(xiàn)糖蛋白的大量合成。它是細(xì)菌糖工程、疫苗開發(fā)和無細(xì)胞糖基化系統(tǒng) (CFGpS) 中的標(biāo)桿級底盤。

BioVector? CLM24 is a genetically optimized Escherichia coli chassis strain specifically designed for protein glycosylation research and production. Derived from the classical $E. coli$ W3110, its defining modification is the complete knockout of the $waaL$ gene ($O$-antigen ligase). By blocking the transfer of endogenous $O$-antigen to lipopolysaccharide (LPS), CLM24 effectively "diverts" lipid-linked oligosaccharide (LLO) substrates to heterologously expressed oligosaccharyltransferases (such as PglB from C. jejuni), enabling high-efficiency glycoprotein synthesis. It is a gold-standard chassis in bacterial glycoengineering, vaccine development, and cell-free glycosylation systems (CFGpS).

BioVector? CLM24 技術(shù)說明書 (Technical Datasheet)

中文版說明書 (Chinese Datasheet)

1. 產(chǎn)品基本信息

• 產(chǎn)品名稱： BioVector? CLM24 糖工程底盤菌株

• 親本菌株：E. coli W3110

• 基因型特征：$W3110, \Delta waaL$。$waaL$ 的缺失防止了 $O$-抗原競爭脂質(zhì)載體 (Und-P)，從而顯著提升糖蛋白產(chǎn)量。

• 主要特性： 具有極高的蛋白質(zhì) $N$-糖基化或 $O$-糖基化修飾效率。

2. 培養(yǎng)條件

• 培養(yǎng)基： BioVector? LB 培養(yǎng)基或 2xYT 培養(yǎng)基。

• 培養(yǎng)溫度： 30 - 37 攝氏度（誘導(dǎo)糖基化時通常建議在 30°C 下以降低包涵體形成）。

• 抗性： 該底盤菌通常無抗性，便于引入多個含不同抗性標(biāo)記的糖基化途徑質(zhì)粒。

• 誘導(dǎo)物： 常用 IPTG 或阿拉伯糖（取決于所載質(zhì)粒的啟動子）。

3. 菌株應(yīng)用

• 生物綴合物疫苗 (Bioconjugates)： 將致病菌的 $O$-抗原結(jié)合到載體蛋白上，開發(fā)針對志賀氏菌、肺炎克雷伯菌等的疫苗。

• 糖蛋白工程： 生產(chǎn)人源化糖基化修飾的治療性蛋白質(zhì)。

• 無細(xì)胞系統(tǒng)： 制備富含糖基化機械的細(xì)胞提取物，用于單罐法 (One-pot) 糖蛋白合成。

4. 注意事項

• LPS 缺陷： 由于 $\Delta waaL$ 導(dǎo)致 LPS 結(jié)構(gòu)不完整（缺失 $O$-抗原），菌株可能對某些表面活性劑或環(huán)境壓力更敏感。

• 驗證方法： 建議通過 SDS-PAGE 和 Western Blot（針對糖鏈的特異性抗體）驗證糖蛋白的修飾比例。

English Datasheet

1. General Product Information

• Product Name: BioVector? CLM24 Glycoengineered Chassis Strain

• Parental Strain:E. coli W3110

• Genotypic Profile:$W3110, \Delta waaL$. The deletion of $waaL$ prevents the competition of $O$-antigen for the lipid carrier (Und-P), significantly increasing the flux toward target glycoproteins.

• Key Characteristics: Optimized for high-efficiency protein $N$-linked or $O$-linked glycosylation.

2. Culture Conditions

• Media: BioVector? LB Broth or 2xYT Broth.

• Temperature: 30°C - 37°C. (Note: 30°C is often preferred during glycosylation induction to minimize protein aggregation).

• Antibiotic Resistance: Typically sensitive to standard antibiotics, allowing for the introduction of multiple plasmids with various resistance markers.

• Inducers: Commonly IPTG or L-arabinose, depending on the plasmid expression system.

3. Applications

• Bioconjugate Vaccine Production: Conjugating pathogenic $O$-antigens onto carrier proteins for vaccines against Shigella, K. pneumoniae, etc.

• Humanized Glycosylation: Engineering bacteria to produce glycoproteins with human-like carbohydrate structures for therapeutics.

• Cell-Free Glyco-platforms: Sourcing lysates enriched with oligosaccharyltransferases (OSTs) and LLOs for rapid, one-pot glycoprotein prototyping.

4. Key Usage Notes

• LPS Integrity: The $\Delta waaL$ phenotype results in "rough" LPS (lacking the $O$-antigen). This may increase sensitivity to certain detergents or environmental stressors compared to wild-type W3110.

• Product Analysis: BioVector? recommends verifying glycosylation efficiency via mass spectrometry or specific lectin-based blotting.

注意 / Note: 在某些前沿研究中，CLM24 也被用于測試抗腫瘤化合物（如吡唑并嘧啶衍生物）。在處理此菌株時，請務(wù)必確認(rèn)您的研究方向是細(xì)菌糖工程還是藥理學(xué)測試。

In some cutting-edge research, the term CLM24 is also associated with antineoplastic pyrazolo[3,4-d]pyrimidine derivatives. Ensure you are utilizing the bacterial chassis strain if your focus is glycoengineering.

BioVector NTCC質(zhì)粒載體菌株細(xì)胞蛋白抗體基因保藏中心

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