Product Features
The human epidermal growth factor receptor HER2 (Her2/neu, ErbB2, or c-erb-b2) is a growth factor receptor that is expressed on many cell types. HER2-proteins are over-expressed in a number of cancer types and assemble in homo- or heterodimers to form functional signaling units. When over-expressed, homo- or heterodimers of HER2 cause increased proliferation, decreased apoptosis, enhanced tumor cell motility and neo-angiogenesis – all being traits strongly associated with tumor formation. Thus, the her2 gene is considered an oncogene.
The Anti-HER2 Affibody® imaging agent is highly specific for the extracellular domain of HER2. It targets HER2-expressing tumors in vivo in SKOV-3 cells in a xenograft model (see Orlova et al., Cancer Res. 2006 Apr 15;66(8):4339-48). The Anti-HER2 imaging agent is a [V1A][D2E] variant of Z00477.
The Anti-HER2 Affibody® imaging agent is modified with a unique C-terminal cysteine for directed single-point chemical modification, facilitating labeling to thiol reactive reagents, such as various fluorescent dyes.
Affibody® products are supplied for research use only and may not be used in a manufacturing process or in manufactured products or in human subjects. The Affibody® products shall in no event be used in medical or clinical applications, unless otherwise expressly stated by Affibody. The purchaser of Affibody® products is solely liable to ensure compliance with any regulatory requirements related to the purchaser’s use of Affibody® products.
Source: Recombinant protein produced in E. coli.
Specificity
Anti-HER2 Affibody® molecule binds to extracellular domain of human epidermal growth factor receptor 2 (HER2). Cross reactivity with other species has not been tested.
MW: 6.9 kDa
Theroetical pI: 8.5
Purity: >95% as determined by RP-HPLC analysis.
Applications
E.g. small animal imaging.Conjugation: The Affibody® molecule contains a unique C-terminal cysteine ideal for directed chemical modifications. However, tail-to-tail dimers are spontaneously generated via a disulfide bridge between the C-terminal cysteines. Prior to coupling via the C-terminal cysteine, the Affibody® molecule needs to be reduced to expose the reactive cysteine residue. Recommended reducing condition is 20 mM DTT at a pH above 7.5 and incubation at room temperature for 2 hours. Remove excess DTT by passage through a desalting column, not by dialysis.
Form: Lyophilized protein. Lyophilized from 10 mM NH₄HCO₃.
Storage
+4°C is recommended for lyophilized protein. For reconstituted protein in physiological buffer, short-term storage at +4°C is recommended. For long-term storage, the protein solution should be aliquoted and then stored at -20°C.
Shipping
At ambient temperature.
Stability
There is no decrease in performance of the reconstituted Affibody® molecule (1 mg/ml in PBS) after 10 repeated freeze and thaw cycles or after storage for 2 weeks at room temperature.
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Reference
Near-infrared photoimmunotherapy targeting EGFR—Shedding new light on glioblastoma treatment
Thomas A. Burley, Justyna Ma˛czynska, Anant Shah, Wojciech Szopa, Kevin J. Harrington, Jessica K.R. Boult, Anna Mrozek-Wilczkiewicz, Maria Vinci, Jeffrey C. Bamber, Wojciech Kaspera and Gabriela Kramer-Marek