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

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


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


Possible Sequences for Secondary Antibody


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


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)


[[../Summary scFv/]]


  • 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.
    • [[../Http://www.sciencedirect.com/science? ob=ArticleURL& udi=B6T1G-44BFHS3-4Y8& user=501045& coverDate=01%2F01%2F1900& fmt=summary& orig=search& qd=1& cdi=4890&view=c& acct=C000022659& version=1& urlVersion=0& userid=501045&md5=31926e4c3e8e19fbea8076163f7dc6a3&ref=full Abstract-EMBASE/]]
    • [[../Http://www.sciencedirect.com/science? ob=ArticleURL& udi=B6WR5-44C90DM-2W6& user=501045& coverDate=01%2F01%2F1900& fmt=summary& orig=search& qd=1& cdi=7009&view=c& acct=C000022659& version=1& urlVersion=0& userid=501045&md5=bb68434e190bccde0798e4939684f6b7&ref=full Abtract-Elsevier BIOBASE/]]
    • [[../Http://www.sciencedirect.com/science? ob=ArticleURL& udi=B6WVB-45DHTFR-4FV& user=501045& coverDate=01%2F01%2F1900& fmt=summary& orig=search& qd=1& cdi=7098&view=c& acct=C000022659& version=1& urlVersion=0& userid=501045&md5=aecd12dab51d29e884ca65f7c41edc99&ref=full Abstract-MEDLINE/]]
    • [[../Http://www.sciencedirect.com/science? ob=RedirectURL& method=outwardLink& partnerName=3& targetURL=http%3A%2F%2Fdx.doi.org%2F10.1038%2F79494& acct=C000022659& version=1& userid=501045&md5=0b0cdc494a012ba14ea1e5d9f685d651 Full Text-CrossRef/]]



Companies checked for antibodies:

eBioscience: no