User:Red Group

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Red Group(an artistic interpretation)

• T/R Section Red Group
• 20.109

Possible Research Topics

1. Fusion of drugs to albumin antibodies to extend in vivo drug half-life
2. Rational design of chimeric restriction enzymes

Research Topic 1

1. Holt LJ, Basran A, Jones K, Chorlton J, Jespers LS, Brewis ND, and Tomlinson IM. Anti-serum albumin domain antibodies for extending the half-lives of short lived drugs. Protein Eng Des Sel. 2008 May;21(5):283-8. DOI:10.1093/protein/gzm067 |

   leave a comment about a paper here

This paper describes how fusing drugs with short half-lives to antibodies specific to albumin, which has a long half-life, extends the drug's half-life without destroying its efficacy.
Another reference is (no PubMed number):
The pharmacokinetics of an albumin-binding Fab can be modulated as a function of affinity for albumin (Nguyen et al.) This paper describes the original creation of albumin-binding antibody-fused species, and the half-life-extending effects of this fusion.

Candidate topics for 20.109:

• Extend work to other drugs
• Test effect on IL-1ra on humans
• Explore other proteins to replace albumin, possibly increasing the effect

Research Topic 2

1. Zhang P, Bao Y, Higgins L, and Xu SY. Rational design of a chimeric endonuclease targeted to NotI recognition site. Protein Eng Des Sel. 2007 Oct;20(10):497-504. DOI:10.1093/protein/gzm049 |

   leave a comment about a paper here

This paper describes the creation of a novel restriction enzyme based off of NotI.

First, the authors isolated a cleavage-deficient variant of NotI restriction enzyme by random mutagenesis. The variant binds to the NotI site but does not cleave it.

Next, the N-terminal cleavage domain from BmrI restriction enzyme was fused to the NotI variant. The BmrI cleaves non-specifically at the N5/N4 site downstream of its recognition sequence.

The chimeric enzyme binds NotI sites when in the presence of high salt and Mg2+ or Ca2+. It cuts outside of the NotI sequence to which it is bound on both sides, essentially removing the site.

"Cleavage site variation has also been observed with natural occurring Type IIG and Type IIS REases. For example, CspCI cut sites can be varied from N10 to N11 at the 50 cut and from N12 to N13 at the 30 cut depending on the adjacent sequence contexts (50-N10 – 11 CAAN5GTGGN12 – 13) (D. Heiter and G. Wilson, unpublished results)."

--Benjamin E. Epstein 00:11, 6 May 2008 (EDT)

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