Human Domain Antibody Library (DAb)
We are pleased distribute the Domain Antibody Phagemid library constructed by Dr Daniel Christ (formally of the MRC Laboratory of Molecular Biology, Cambridge UK and currently based at the Garvan Institute, Sydney, Australia).
This library is based on a VH framework (V3-23/D47). Diversity was introduced into the antigen binding domains by PCR mutagenesis into CDR1, CDR2 and CDR3. There are many benefits to this library.
- Produced binders to the model antigens refold without major loss of antigen-binding activity upon cooling.
- Antibodies selected maintain antigen-binding activity after up to 25 cycles of heat denaturation, rivalling the heat resistance of thermophilic protein such as Taq polymerase.
- Repertoire withstands heat-induced aggregation on phage.
- These benefits offer a wide range of diagnostic and therapeutic research applications and can further maximise the number of binders to your antigen of interest.
The use of the resource is limited to research purposes. The collection is subject to the following MTA and our clone terms and conditions
This library is supplied as a single kit containing:
- Phagemid antibody library (~3x109)
- Helper phage KM13
- Host strain TG1Tr
- Optimised antibody expression strain HB2151
- Controls: anti-beta-galactosidase & anti-bovine ubiquitin antibodies
- detailed protocol
This artificial library of antibodies can be used to derive binders to almost any target molecule using phage display and selection. These binders can be used for the same applications as conventional monoclonal antibodies (ELISA, Western blotting, FACS, immunohistochemistry), but can be isolated in a fraction of the time and without the need for animal immunisation.
Phage-antibody libraries have been used successfully in hundreds of molecular biology labs worldwide to derive highly specific antibody reagents to a wide range of different proteins, peptides or small molecule compounds.
||Human Domain Antibody Constructs (DAb)
The KM13 Helper Phage are included in the DAb library, however they can be ordered as a separate item below:
||KM13 Helper Phage
The use of the resource is limited to research purposes. The collection is subject to the following MTA and our clone terms and conditions.
This library is based on a VH framework (V3-23/D47). Diversity was introduced into the antigen binding domains by PCR mutagenesis into CDR1, CDR2 and CDR3.
Library: Human Domain Antibody Library (DAb)
Source Species: Homo sapiens;
Host Species: E. coli; Host Strain: TG1
Growth Conditions medium: M9 minimal medium, 2xTY medium
Growth Conditions antibiotic: Amp
The sequence/map of pR2 can be viewed in pdf or fasta formats.
Please note: The sequence contains a ‘example’ sequence for the VH which is called DummyVH, and is marked in red in the pdf file.
Unfortunately there are no primers listed in the library protocol or in the publications for the DAb library, however there is an LMB3 (5′-CAGGAAACAGCTATGAC-3′) site upstream of the insert and an fdSeq1 site (5′-GAATTTTCTGTATGAGG-3′) within the Gene III sequence.
Dass, S.A., Norazmi, M.N., Dominguez, A.A., San Miguel, M.E.S.G. and Tye, G.J., 2018. Generation of a T cell receptor (TCR)-like single domain antibody (sDAb) against a Mycobacterium Tuberculosis (Mtb) heat shock protein (HSP) 16kDa antigen presented by Human Leukocyte Antigen (HLA)-A* 02. Molecular immunology, 101, pp.189-196.
Niccheri, F., Real-Fernàndez, F., Ramazzotti, M., Lolli, F., Rossi, G., Rovero, P. and Degl'Innocenti, D. (2014),Human recombinant domain antibodies against multiple sclerosis antigenic peptide CSF114(Glc).J. Mol. Recognit., 27: 618–626. doi:10.1002/jmr.2386>
Lee CM, Iorno N, Sierro F, Christ D.
Selection of human antibody fragments by phage display.
Nat Protoc. 2007;2(11):3001-8.
Dudgeon K, Famm K, Christ D.
Sequence determinants of protein aggregation in human VH domains.
Protein Eng Des Sel. 2008 Oct 28.
Famm K, Hansen L, Christ D, Winter G.
Thermodynamically stable aggregation-resistant antibody domains through directed evolution.
J Mol Biol. 2008 Feb 29;376(4):926-31.
Christ D, Famm K, Winter G.
Repertoires of aggregation-resistant human antibody domains.
Protein Eng Des Sel. 2007 Aug;20(8):413-6.
Christ D, Famm K, Winter G.
Tapping diversity lost in transformations--in vitro amplification of ligation reactions.
Nucleic Acids Res. 2006;34(16):e108.
For further information and prices please contact us or call +44 (0)115 973 9012