Difference between revisions of "Part:BBa K929102"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K929102 short</partinfo> | <partinfo>BBa_K929102 short</partinfo> | ||
| − | + | <p style="background-color: rgb(238, 221, 130); font-weight: bold;">General information</p> | |
{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right" | {| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right" | ||
| − | ! colspan="2" style="background: | + | ! colspan="2" style="background:rgb(240, 20, 70);"| Anti-GFP Nanobody with IgG kappa signal peptide and IgG1 Fc domain |
|- | |- | ||
! colspan="2"|[[Image:UP12_BBa_K929102.png|300px]] | ! colspan="2"|[[Image:UP12_BBa_K929102.png|300px]] | ||
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|'''Requirement''' | |'''Requirement''' | ||
| − | |pSB1C3 | + | |pSB1C3 |
|- | |- | ||
|'''Source''' | |'''Source''' | ||
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|[http://2012.igem.org/Team:Potsdam_Bioware Potsdam_Bioware2012] | |[http://2012.igem.org/Team:Potsdam_Bioware Potsdam_Bioware2012] | ||
|} | |} | ||
| − | + | ||
[[Image:UP12_BBa_K929102_vector.png|left|thumb|400px]]<br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br> | [[Image:UP12_BBa_K929102_vector.png|left|thumb|400px]]<br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br> | ||
<br> | <br> | ||
<br> | <br> | ||
| − | This part represents an antibody unit composed of a IgG kappa signal peptide, an anti-GFP nanobody and an IgG1 Fc domain.<br> | + | This part represents an antibody unit composed of a IgG kappa signal peptide, an anti-GFP nanobody and an IgG1 Fc domain. <br> |
<br> | <br> | ||
| − | Signal peptide IgG kappa:<br> | + | '''Signal peptide IgG kappa:'''<br> |
| − | The human signal peptide of the immunoglobulin G kappa variable chain I has 22 aa and is responsible for the translocation of the fusion protein to the membrane. | + | The human signal peptide of the immunoglobulin G kappa variable chain I has 22 aa and is responsible for the translocation of the fusion protein to the membrane (UniProt: P01601). |
<br> | <br> | ||
<br> | <br> | ||
| − | Anti-GFP nanobody:<br> | + | '''Anti-GFP nanobody:'''<br> |
| − | The green fluorescent protein (GFP)-nanobody is a single-chain VHH antibody domain developed with specific binding activity against GFP and shows a Kd value of 1.4 nM. Its CDR3 loop is very short and has significantly fewer contacts with the GFP ligand compared to other nanobodies. Furthermore, the shortness of this CDR3 loop leads to the exposure of the framework 2 region, which has a major contribution to the binding with GFP. | + | The green fluorescent protein (GFP)-nanobody (PDB: 3OGO) is a single-chain VHH antibody domain developed with specific binding activity against GFP and shows a Kd value of 1.4 nM. Its CDR3 loop is very short and has significantly fewer contacts with the GFP ligand compared to other nanobodies. Furthermore, the shortness of this CDR3 loop leads to the exposure of the framework 2 region, which has a major contribution to the binding with GFP. |
<br> | <br> | ||
<br> | <br> | ||
| − | Fc region:<br> | + | '''Fc region:'''<br> |
| − | A human immunoglobulin gamma-1 heavy chain constant region (IGHG1) was added to the construct and enables the direct interaction of the antibody unit with the Fc receptor and complement proteins. | + | A human immunoglobulin gamma-1 heavy chain constant region (IGHG1) was added to the construct and enables the direct interaction of the antibody unit with the Fc receptor and complement proteins ( UniProt: P01857). |
<br> | <br> | ||
| + | <p style="background-color: rgb(238, 221, 130); font-weight: bold;">Characterization</p> | ||
| + | For characterization and further applications see BBa_K929107. | ||
| − | |||
===Usage and Biology=== | ===Usage and Biology=== | ||
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<partinfo>BBa_K929102 parameters</partinfo> | <partinfo>BBa_K929102 parameters</partinfo> | ||
<!-- --> | <!-- --> | ||
| + | |||
| + | <h2><b>Contribution of 2022 Sydney iGEM</b></h2> | ||
| + | We have modelled the binding of this nanobody to sfGFP and fuGFP. | ||
| + | [[File:modelling-sf-h.gif|centre|thumb|700px| modelling of Nanobody h (3OGO) to sfGFP]] | ||
| + | [[File:modelling-fu-h.gif|centre|thumb|700px| modelling of Nanobody h (3OGO) to sfGFP]] | ||
Latest revision as of 09:58, 12 October 2022
Anti-GFP Nanobody with IgG kappa signal peptide and IgG1 Fc domain
General information
| Anti-GFP Nanobody with IgG kappa signal peptide and IgG1 Fc domain | |
|---|---|
| |
| BioBrick Nr. | BBa_929102 |
| RFC standard | RFC 25 |
| Requirement | pSB1C3 |
| Source | Gene synthesis by GeneArt |
| Submitted by | [http://2012.igem.org/Team:Potsdam_Bioware Potsdam_Bioware2012] |
This part represents an antibody unit composed of a IgG kappa signal peptide, an anti-GFP nanobody and an IgG1 Fc domain.
Signal peptide IgG kappa:
The human signal peptide of the immunoglobulin G kappa variable chain I has 22 aa and is responsible for the translocation of the fusion protein to the membrane (UniProt: P01601).
Anti-GFP nanobody:
The green fluorescent protein (GFP)-nanobody (PDB: 3OGO) is a single-chain VHH antibody domain developed with specific binding activity against GFP and shows a Kd value of 1.4 nM. Its CDR3 loop is very short and has significantly fewer contacts with the GFP ligand compared to other nanobodies. Furthermore, the shortness of this CDR3 loop leads to the exposure of the framework 2 region, which has a major contribution to the binding with GFP.
Fc region:
A human immunoglobulin gamma-1 heavy chain constant region (IGHG1) was added to the construct and enables the direct interaction of the antibody unit with the Fc receptor and complement proteins ( UniProt: P01857).
Characterization
For characterization and further applications see BBa_K929107.
Usage and Biology
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Contribution of 2022 Sydney iGEM
We have modelled the binding of this nanobody to sfGFP and fuGFP.
