Difference between revisions of "Part:BBa K929102"

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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

Contribution of 2022 Sydney iGEM

We have modelled the binding of this nanobody to sfGFP and fuGFP.

modelling of Nanobody h (3OGO) to sfGFP

@@ Line 3: / Line 3: @@
 <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"
-! colspan="2" style="background:rgb(240, 20, 70);"|[https://parts.igem.org/Part:BBa_K929002 Anti-GFP Nanobody with IgG kappa signal peptide and IgG1 Fc domain]
+! 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]]
@@ Line 14: / Line 14: @@
 |-
 |'''Requirement'''
-|pSB1C3<br>
+|pSB1C3
 |-
 |'''Source'''
@@ Line 22: / Line 22: @@
 |[http://2012.igem.org/Team:Potsdam_Bioware Potsdam_Bioware2012]
 |}
-<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>
-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>
-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>
-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>
-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>
+<p style="background-color: rgb(238, 221, 130); font-weight: bold;">Characterization</p>
+For characterization and further applications see BBa_K929107.
-<!-- Add more about the biology of this part here
 ===Usage and Biology===
@@ Line 52: / Line 53: @@
 <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]]