Part Collection
Our part collection contains a variety of nanobody-based composite parts including secretion tags, and the whole secretion system construct. Nanobody parts are derived from 3 single nanobodies obtained from the patent mentioned in the registry. 1 Combination of these 3 nanobodies resulted in 9 nanobody constructs with different properties which we characterized with western blots, ELISA and immunoassays on human monocytes. Composite parts of the different nanobodies together with the inducible pBAD promoter and HlyA-tag for secretion are included. Together with the construct that contains the secretion system, a whole system that allows the secretion of anti-TNFα nanobodies can be built and transformed into any suitable strain that expresses TolC (needed for a functional secretion system). Exchanging certain BioBricks, such as the nanobodies, allows future iGEM teams to adjust this collection towards their needs, for example to use the system to secrete other proteins of interest.
Here is an overview of all the parts that belong to the collection.
| Name | Type | Description | Designers | Length | Picture |
|---|---|---|---|---|---|
| BBa_K4387987 | Secretion system | Hemolysin A secretion system for E. coli | Nathalie Weibel | 3734 |
|
| BBa_K4387986 | Monovalent nanobody expression | Monovalent nanobody expression with the HlyA secretion signal (VHH#2B) | Nathalie Weibel | 2421 |
|
| BBa_K4387985 | Monovalent nanobody expression | Monovalent nanobody expression with the HlyA secretion signal (VHH#3E) | Nathalie Weibel | 2439 |
|
| BBa_K4387984 | Monovalent nanobody expression | Monovalent nanobody expression with the HlyA secretion signal (VHH#12B) | Nathalie Weibel | 2421 |
|
| BBa_K4387983 | Bivalent nanobody expression | Bivalent nanobody expression with the HlyA secretion signal (biv. VHH#2B) | Nathalie Weibel | 2826 |
|
| BBa_K4387982 | Bivalent nanobody expression | Bivalent nanobody expression with the HlyA secretion signal (biv. VHH#3E) | Nathalie Weibel | 2862 |
|
| BBa_K4387981 | Bivalent nanobody expression | Bivalent nanobody expression with the HlyA secretion signal (VHH#3E + VHH#2B) | Nathalie Weibel | 2844 |
|
| BBa_K4387980 | Bivalent nanobody expression | Bivalent nanobody expression with the HlyA secretion signal (VHH#2B + VHH#12B) | Nathalie Weibel | 2826 |
|
| BBa_K4387979 | Bivalent nanobody expression | Bivalent nanobody expression with the HlyA secretion signal (VHH#3E + VHH#12B) | Nathalie Weibel | 2844 |
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References:
- Silence, Karen, Lauwereys, Marc, De Haard, Hans, et al. "Single domain antibodies directed against tumour necrosis factor-alpha and uses therefor", Int. Publication Number: WO 2004/041862 A2, 21 May 2004