IGEM:MIT/2005/Antibodies

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Contents

TO DO LIST

1) Obtain plasmid with anti-rabbit scFV from authors of the SCFV antibody Piezoimmunosensors paper

Zeng@oakland.edu
Ask
NCBI sequence different from sequence in paper
conserved region
get clone on plasmid

2) Determine and order fluorescein - tagged DNA for use in determining dimerization distance

3) Blast and check the "low complexity" region of the light chain in the scFV

confirm the match to rice firus is a fluke!

4) Check 3D structure of antibodies (ask Natalie and Kate)

5) Additional info: swiss-prot

Goals

  • Find an antigen and its antibodies
  • Sequence of the fusing antibody
  • Proper distance between epitopes

Resources

Possible Sequences for Secondary Antibody

Anti-rabbit

Sequence: Sequence of synthetic construct anti-rabbit IgG scFV mRNA, partial cds
Paper: scFv Antibody Piezoimmunosensors
Summary: Summary scFvSensor

Anti-mouse

ScFVs: single chain fragment variable (we shouldn't have to engineer these too much)
Sequence anti-mouse1: Synthetic construct clone H2 anti-mouse L1 scFv antibody mRNA, partial cds.
Sequence anti-mouse2: Synthetic construct clone G6 anti-mouse L1 scFv antibody mRNA, partial cds.
Paper covering anti-mouse1 and 2: Single chain Fv antibodies against neural cell adhesion molecule L1 trigger L1 functions in cultured neurons
Summary of anti-mouse1 and 2 paper: Summary anti-mouse 1 and 2
Sequence of anti-mouse3: Synthetic construct clone C12 anti-mouse CHL1 scFv antibody mRNA, partial cds.
Sequence of amti-mouse4: Synthetic construct clone 6C2 anti-mouse CHL1 scFv antibody mRNA, partial cds.
Paper covering anti-mouse3 and 4: Single chain variable fragment antibodies against the neural adhesion molecule CHL1 (close homolog of L1) enhance neurite outgrowth
Summary of anti-mouse3 and 4: Summary anti-mouse 3 and 4
Light and Heavy Chains (we will need to engineer a linker sequence for these sequences)

Papers

Summary scFv


Books

  • Sequences of Proteins of Immunological Interest, Kabat. Call Number: QD431.S59 1991.
  • Antibody Engineering: A practical approach, McCafferty et al. 1996

Questions, Comments, Concerns, Suggestions

  • R.M. de Wildt, C.R. Mundy, B.D. Gorick and I.M. Tomlinson, Antibody arrays for high-throughput screening of antibody–antigen interactions. Nat. Biotechnol. 18 (2000), pp. 989–994.

Diagrams

Misc

Companies checked for antibodies:

eBioscience: no

Anit-Florouscine

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