BioVector® CLM24 糖工程大肠杆菌底盘菌株 / BioVector® CLM24 Glycoengineered E. coli Chassis Strain

通用定义 / General Definition:

BioVector® CLM24 是一种经过基因工程高度优化的大肠杆菌 (Escherichia coli) 底盘菌株，专门用于蛋白质糖基化 (Protein Glycosylation) 的研究与生产。该菌株源自经典的 $E. coli$ W3110，其核心改造是彻底敲除了 $waaL$ 基因（$O$-抗原连接酶基因）。通过阻断内源性 $O$-抗原向脂多糖 (LPS) 的转移，CLM24 能够将脂质载体上的聚糖底物高效地“让渡”给异源表达的寡糖转移酶（如来自空肠弯曲杆菌的 PglB），从而实现糖蛋白的大量合成。它是细菌糖工程、疫苗开发和无细胞糖基化系统 (CFGpS) 中的标杆级底盘。

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 技术说明书 (Technical Datasheet)

中文版说明书 (Chinese Datasheet)

1. 产品基本信息

• 产品名称： BioVector® CLM24 糖工程底盘菌株

• 亲本菌株：E. coli W3110

• 基因型特征：$W3110, \Delta waaL$。$waaL$ 的缺失防止了 $O$-抗原竞争脂质载体 (Und-P)，从而显著提升糖蛋白产量。

• 主要特性： 具有极高的蛋白质 $N$-糖基化或 $O$-糖基化修饰效率。

2. 培养条件

• 培养基： BioVector® LB 培养基或 2xYT 培养基。

• 培养温度： 30 - 37 摄氏度（诱导糖基化时通常建议在 30°C 下以降低包涵体形成）。

• 抗性： 该底盘菌通常无抗性，便于引入多个含不同抗性标记的糖基化途径质粒。

• 诱导物： 常用 IPTG 或阿拉伯糖（取决于所载质粒的启动子）。

3. 菌株应用

• 生物缀合物疫苗 (Bioconjugates)： 将致病菌的 $O$-抗原结合到载体蛋白上，开发针对志贺氏菌、肺炎克雷伯菌等的疫苗。

• 糖蛋白工程： 生产人源化糖基化修饰的治疗性蛋白质。

• 无细胞系统： 制备富含糖基化机械的细胞提取物，用于单罐法 (One-pot) 糖蛋白合成。

4. 注意事项

• LPS 缺陷： 由于 $\Delta waaL$ 导致 LPS 结构不完整（缺失 $O$-抗原），菌株可能对某些表面活性剂或环境压力更敏感。

• 验证方法： 建议通过 SDS-PAGE 和 Western Blot（针对糖链的特异性抗体）验证糖蛋白的修饰比例。

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 也被用于测试抗肿瘤化合物（如吡唑并嘧啶衍生物）。在处理此菌株时，请务必确认您的研究方向是细菌糖工程还是药理学测试。

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质粒载体菌株细胞蛋白抗体基因保藏中心

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