I am going to make plasmid construction, contain certain scFv (small protein fragment approximately 30-100 kD) gene and express it by means of a common expression E.coli strain.
then it will be conjugated on Gold nano-particle surface for targeting purpose.
what factors must be considered in nano particles-protein conjugation???
Usually disulfide bonds are used for conjugation of molecule to gold nano-particle. if your protein has a cysteine, it is possible.
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First thanks for response .
It is possible to insert cysteine at the end of scFv (N-terminal).
One aa is enough?
See attached [1] and references cintained within for discussion on factors leading to the stablity of gold - antibody conjugates: E.G.:
Passive absorption onto gold particles is often directed by the isoelectric point, pI of the protein, though this is not allways so and can be directed by say an electro-negative patch on the protein surface. The particle surface charge, as reflected in the particle zeta-potential, also plays a role in protein - particle conugate stability / absorption. Thus the conditions which promote stable conjugate formation often includes pH and concentration ratio of particles to protein
You may also wish to dip into DLVO theory [2].
If, however, your going to produce your own scFv fragment protein, then add in a cysteine [3] to help direct the scFv absorption and orientation onto the gold particles.
- John.
[1] Chemical Modification of Antibody Enables the Formation of Stable Antibody-Gold Nanoparticle Conjugates for Biosensing, Analyst. 2017 Nov 20;142(23):4456-4467
[2] https://en.wikipedia.org/wiki/DLVO_theory
[3] Single chain fragment variable recombinant antibody functionalized gold nanoparticles for a highly sensitive colorimetric immunoassay, Biosensors and Bioelectronics 24 (2009) 2853–2857
Because your molecule is small, adding an amino acid may be enough. But in general, if the number of disulfide bonds is higher, you have a better conjugation of molecule to gold nano-particle.
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