Product Features
The Epidermal Growth Factor Receptor (EGFR, ErbB) is a cell surface glycoprotein of approximately 135 kDa (unglycosylated). There are several alternatively spliced forms of EGFR including a secreted soluble form. The EFGR belongs to the family tyrosine kinase receptors which are characterized by an extracellular ligand-binding domain, a single transmembrane and an intracellular domain responsible for transducing the signal. The receptor dimerizes upon binding to EGFR and the transduced signal stimulates cell growth and differentiation. The EGFR is over expressed or mutated in many types of cancers and the receptor as well as the down stream signaling molecules are subjects to several cancer therapeutic interventions.
The Anti-EGFR Affibody® molecule is specific for the extra cellular domain of EGFR.
The Anti-EGFR 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-EGFR Affibody® Imaging Agent binds to the extracellular domain of human epidermal growth factor receptor (EGFR, ErbB). Cross reactivity with other species has not been tested.
Mw: 6.7 kDa
Theoretical pI: 4.6
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
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. For longer term storage the protein solution should first be aliquoted and stored frozen at −20 °C.
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References
Near-infrared photoimmunotherapy targeting EGFR—Shedding new light on glioblastoma treatment
Thomas A. Burley, Justyna Maczynska , 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
Affibody-Based PET Imaging to Guide EGFR-Targeted Cancer Therapy in Head and Neck Squamous Cell Cancer Models
Thomas A. Burley, Chiara Da Pieve, Carlos D. Martins, Daniela M. Ciobota, Louis Allott1, Wim J.G Oyen, Kevin J. Harrington, Graham Smith, and Gabriela Kramer-Marek