Show simple item record

dc.creatorJackson, Michael Gary
dc.date.accessioned2012-06-07T22:45:03Z
dc.date.available2012-06-07T22:45:03Z
dc.date.created1996
dc.date.issued1996
dc.identifier.urihttps://hdl.handle.net/1969.1/ETD-TAMU-1996-THESIS-J336
dc.descriptionDue to the character of the original source materials and the nature of batch digitization, quality control issues may be present in this document. Please report any quality issues you encounter to digital@library.tamu.edu, referencing the URI of the item.en
dc.descriptionIncludes bibliographical references.en
dc.descriptionIssued also on microfiche from Lange Micrographics.en
dc.description.abstractSingle chain antibody (scfv NC6.8) was generated from the IgG (2b,k) parent mAb cell line NC6.8, a mAb that binds a super-potent trisubstituted guanidino sweetener, N-(p-cyanophenyl)-N'(diphenylmethyl)guanidine acetic acid. The plasmid construct was cloned from the VH and VL MRNA isolated from the parent mAb cell line. Expression was induced in transformed E. coli and scfv was localized in inclusion bodies. ScFv was isolated and solubilized in 6 M Gdn-HCI. Three different methods of affinity purification were compared: 1) anti c-myc mAb for a c-myc-scfv NC6.8 fusion construct, 2) metal chelation for a histidine hexamer-scfv NC6.8 fusion construct, and 3) polyclonal anti-scfv NC 6.8 antibody, for either of the above fusion constructs. The polyclonal sheep antibody to the scfv was prepared using the NC6.8 Fab as an antigen. This antisera was then absorbed to remove polyclonal anti-lg(CH1+CL) activity prior to affinity purification with a NC6.8 IgG column. Antic-myc mabs (9ElO and CT14 cell lines) that react with residues 408439 of the c-myc peptide were grown as ascites; purified IgG was coupled to Affi-gel-10 resin. Identification of purified scfv was accomplished by immunoblotting and ELISA. Analysis using circular dichroism revealed a predominantly D-character of the pure scfv. Determinations of total yield, purity, and activity were compared using different affinity purification techniques.en
dc.format.mediumelectronicen
dc.format.mimetypeapplication/pdf
dc.language.isoen_US
dc.publisherTexas A&M University
dc.rightsThis thesis was part of a retrospective digitization project authorized by the Texas A&M University Libraries in 2008. Copyright remains vested with the author(s). It is the user's responsibility to secure permission from the copyright holder(s) for re-use of the work beyond the provision of Fair Use.en
dc.subjectmicrobiology.en
dc.subjectMajor microbiology.en
dc.titleComparative affinity purification of single chain antibody NC6.8en
dc.typeThesisen
thesis.degree.disciplinemicrobiologyen
thesis.degree.nameM.S.en
thesis.degree.levelMastersen
dc.type.genrethesisen
dc.type.materialtexten
dc.format.digitalOriginreformatted digitalen


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record

This item and its contents are restricted. If this is your thesis or dissertation, you can make it open-access. This will allow all visitors to view the contents of the thesis.

Request Open Access