1ap2

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{{STRUCTURE_1ap2| PDB=1ap2 | SCENE= }}
{{STRUCTURE_1ap2| PDB=1ap2 | SCENE= }}
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'''SINGLE CHAIN FV OF C219'''
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===SINGLE CHAIN FV OF C219===
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==Overview==
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A construct encoding a single chain variable fragment of the anti-P-glycoprotein monoclonal antibody C219 was made by combining the coding sequences for the heavy and light chain variable domains with a sequence encoding the flexible linker (GGGGS)3, an OmpA signal sequence, a c-myc identification tag, and a five-histidine purification tag. The construct was expressed in Escherichia coli and purified from the periplasmic fraction using a nickel chelate column and ion exchange chromatography. Three-step Western blot analysis showed that the construct retains binding affinity for P-glycoprotein. Crystals of 1.0 x 0.2 x 0.2 mm were grown in 100 mM citrate, pH 4.5, 21% polyethylene glycol 6000 in the presence of low concentrations of subtilisin, resulting in proteolytic removal of the linker and purification tags. The structure was solved to a resolution of 2.4 A with an R factor of 20.6, an Rfree of 28.5, and good stereochemistry. This result could lead to a clinically useful product based on antibody C219 for the diagnosis of P-glycoprotein-mediated multidrug resistance. The molecule will also be useful in biophysical studies of functional domains of P-glycoprotein, as well as studies of the intact molecule.
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{{ABSTRACT_PUBMED_9368049}}
==About this Structure==
==About this Structure==
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[[Category: P-glycoprotein]]
[[Category: P-glycoprotein]]
[[Category: Single chain fv]]
[[Category: Single chain fv]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Fri May 2 10:32:21 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Jun 30 17:19:56 2008''

Revision as of 14:19, 30 June 2008

Image:1ap2.png

Template:STRUCTURE 1ap2

SINGLE CHAIN FV OF C219

Template:ABSTRACT PUBMED 9368049

About this Structure

1AP2 is a Protein complex structure of sequences from Mus musculus. Full crystallographic information is available from OCA.

Reference

A single chain Fv fragment of P-glycoprotein-specific monoclonal antibody C219. Design, expression, and crystal structure at 2.4 A resolution., Hoedemaeker FJ, Signorelli T, Johns K, Kuntz DA, Rose DR, J Biol Chem. 1997 Nov 21;272(47):29784-9. PMID:9368049

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