Hi, I am trying to make protein adsorbed on polymer particles.
I incubated protein and polymer solution in 70 °C because I thought
thermal denaturation can increase the amount of adsorption and I got some white aggregated proteins in the sample.
However, I am trying to measure the concentration of supernatant to identify the adsorbed amount of protein.
Therefore, I am trying to dissolve the aggregated denatured protein again by sonication.
The question is, does this sonication process will desorb the adsorbed protein on polymer particles?
Or is it impossible to solubilize again thermally denatured protein by sonication, and do I have to
redesign the experiment procedure??
What is the surface charge of the particles at different pHs?
Instead of thermal denaturation, I recommend adjustment of the solution pH at/or close to the pI value of the protein. Otherwise, set the pH to a value that helps the electrostatic adsorption. If the particles are inert, you may need surface functionalization of the particles. The sonication would desorb any adsorbed protein.
Please c.f.
Article High-Capacity Hierarchically Imprinted Polymer Beads for Pro...
Article Hierarchically structured protein-based hollow-nanospheres f...
Ca Dr. Gaurav Bhambri
You have denatured the protein by taking it to 700C - rather like cooking an egg. The temperature in a collapsing sonication bubble is around 5000 K and, although the bubble is small, one manifestation of the working of sonication is an increase in temperature which will also cook the protein. See this webinar (registration required) Slide 43:
Ultrasound, cavitation, and the singing kettle
https://www.malvernpanalytical.com/en/learn/events-and-training/webinars/W130619UltrasoundCavitationSingingKettle.html
I agree with the previous poster in that change of pH or by adsorption of one species (e.g. phosphate) on the polymer followed by adsorption of the protein. Proteins are often zwitterionic (2 opposite charges on the same molecule) so some experimentation will be needed.
Also check https://www.sciencedirect.com/topics/chemistry/adsorption-of-protein
Thanks for your's gracious participation Chinaza Godswill Awuchi
Kindly check on this paper
Article Protein Adsorption on Various Plasma-Treated Polyethylene Te...
Article High-Capacity Hierarchically Imprinted Polymer Beads for Pro...
Article Hierarchically structured protein-based hollow-nanospheres f...
bovine serum albumin (BSA) hollow-nano-spheres (HNS) were synthesized as nano-carriers for paclitaxel (PTX) as model drug. Hierarchically, first, the exterior surface of silica nanoparticles was modified with (3-aminopropyl)triethoxysilane (APTES) and then with glutaraldehyde (GA). Then, BSA at alkaline pH was adsorbed on the surface of the modified nanoparticles and further crosslinked with GA
https://www.researchgate.net/deref/https%3A%2F%2Fwww.sciencedirect.com%2Ftopics%2Fchemistry%2Fadsorption-of-protein
https://www.researchgate.net/deref/https%3A%2F%2Fwww.sciencedirect.com%2Ftopics%2Fchemistry%2Fadsorption-of-protein
https://www.researchgate.net/deref/https%3A%2F%2Fwww.sciencedirect.com%2Ftopics%2Fchemistry%2Fadsorption-of-protein
https://www.researchgate.net/deref/https%3A%2F%2Fedepot.wur.nl%2F121742
https://www.researchgate.net/deref/https%3A%2F%2Fedepot.wur.nl%2F121742
Actually, The influence of protein stability on the adsorption and desorption behavior to surfaces with fundamentally different properties (negatively charged, positively charged, hydrophilic, and hydrophobic) was examined by surface plasmon resonance measurements. Three engineered variants of human carbonic anhydrase II were used that have unchanged surface properties but large differences in stability. The orientation and conformational state of the adsorbed protein could be elucidated by taking all of the following properties of the protein variants into account: stability, unfolding, adsorption, and desorption behavior. Regardless of the nature of the surface, there were correlation between (i) the protein stability and kinetics of adsorption, with an increased amplitude of the first kinetic phase of adsorption with increasing stability; (ii) the protein stability and the extent of maximally adsorbed protein to the actual surface, with an increased amount of adsorbed protein with increasing stability; (iii) the protein stability and the amount of protein desorbed upon washing with buffer
I think In this case, the surface has to be engineered to avoid or govern the adsorption process (6 – 8). The second area is when a specific protein comes into contact with a large number of surfaces, e.g. in the downstream processing, polishing, storage, and final use of proteins produced for biotechnological or medical applications
May be In this case, it is unlikely that all of the surfaces (glass, plastic, stainless steel, rubber, and so forth) that a specific protein is exposed to during production and use can be made resistant to protein adsorption, i.e. it could be more feasible to engineer the protein to make it less prone to adsorb onto surfaces. Although this case has not been given as much attention, it is interesting to note that the downstream processing of therapeutic antibodies accounts for an estimated 50 – 80% of the total manufacturing cost
https://www.researchgate.net/post/Sonication_will_desorb_the_adsorbed_protein#:~:text=May%20be%20In,total%20manufacturing%20cost
https://www.researchgate.net/post/Sonication_will_desorb_the_adsorbed_protein#:~:text=May%20be%20In,total%20manufacturing%20cost
Contact Angle Goniometry—Contact angles were measured with a Rame´-Hart NRL 100 goniometer with no control of ambient humidity. As contact liquids, freshly deionized water from a Milli-Q unit and hexadecane (Merck) were used, respectively. Two separate measurements were performed on each plate.
https://www.researchgate.net/post/Sonication_will_desorb_the_adsorbed_protein#:~:text=Contact%20Angle%20Goniometry%E2%80%94Contact%20angles%20were%20measured%20with%20a%20Rame%C2%B4-Hart%20NRL%20100%20goniometer%20with%20no%20control%20of%20ambient%20humidity.%20As%20contact%20liquids%2C%20freshly%20deionized%20water%20from%20a%20Milli-Q%20unit%20and%20hexadecane%20(Merck)%20were%20used%2C%20respectively.%20Two%20separate%20measurements%20were%20performed%20on%20each%20plate.
https://www.researchgate.net/post/Sonication_will_desorb_the_adsorbed_protein#:~:text=Contact%20Angle%20Goniometry%E2%80%94Contact%20angles%20were%20measured%20with%20a%20Rame%C2%B4-Hart%20NRL%20100%20goniometer%20with%20no%20control%20of%20ambient%20humidity.%20As%20contact%20liquids%2C%20freshly%20deionized%20water%20from%20a%20Milli-Q%20unit%20and%20hexadecane%20(Merck)%20were%20used%2C%20respectively.%20Two%20separate%20measurements%20were%20performed%20on%20each%20plate.
https://www.researchgate.net/post/Sonication_will_desorb_the_adsorbed_protein#:~:text=May%20be%20In,total%20manufacturing%20cost
https://www.researchgate.net/post/Sonication_will_desorb_the_adsorbed_protein#:~:text=May%20be%20In,total%20manufacturing%20cost
The influence of protein stability on the adsorption and desorption behavior to surfaces with fundamentally different properties (negatively charged, positively charged, hydrophilic, and hydrophobic) was examined by surface plasmon resonance measurements. Three engineered variants of human carbonic anhydrase II were used that have unchanged surface properties but large differences in stability. The orientation and conformational state of the adsorbed protein could be elucidated by taking all of the following properties of the protein variants into account
https://www.researchgate.net/deref/https%3A%2F%2Fwww.jbc.org%2Farticle%2FS0021-9258(20)61367-9%2Ffulltext
https://www.researchgate.net/deref/https%3A%2F%2Fedepot.wur.nl%2F121742
https://www.researchgate.net/deref/https%3A%2F%2Fedepot.wur.nl%2F121742
Most probabaly, Three engineered variants of human carbonic anhydrase II were used that have unchanged surface properties but large differences in stability. The orientation and conformational state of the adsorbed protein could be elucidated by taking all of the following properties of the protein variants into account
Whereas, As contact liquids, freshly deionized water from a Milli-Q unit and hexadecane (Merck) were used, respectively. Two separate measurements were performed on each plate.
Actually, The orientation and conformational state of the adsorbed protein could be elucidated by taking all of the following properties of the protein variants into account
Three engineered variants of human carbonic anhydrase II were used that have unchanged surface properties but large differences in stability
https://www.researchgate.net/post/Sonication_will_desorb_the_adsorbed_protein#:~:text=May%20be%20In,total%20manufacturing%20cost
Thanks a lot for gracious participation respected Dr Sushma Tayal
Thanks a lot for gracious participation respected Dr keshav Bhambri
Actually, The orientation and conformational state of the adsorbed protein could be elucidated by taking all of the following properties of the protein variants into account
Surfaces that resist the adsorption of proteins are essential for certain applications in biomaterials and for research in biosurface chemistry. SAMs presenting oligomers of ethylene glycol, prepared using the al-kanethiols HS(CH2)11(OCH2CH2)n OH(EGn), resist the adsorption of proteins to surfaces [38,39].
But , adsorption of proteins are essential for certain applications in biomaterials and for research in biosurface chemistry. SAMs presenting oligomers of ethylene glycol, prepared using the al-kanethiols HS(CH2)11(OCH2CH2)n OH(EGn), resist the adsorption of proteins to surfaces [38,39].
The EGn-terminated SAMs are not unique in their ability to resist the adsorption of proteins. SAMs presenting tripropylene sulfoxide groups also prevent the non-specific adsorption of proteins [40].
The adsorption of proteins on synthetic surfaces, such as polystyrene latex and glass, was studied as early as 1956 (26) although its popular application to immunoassay dates to Catt and Tregear
The adsorption of proteins on synthetic surfaces, such as polystyrene latex and glass, was studied as early as 1956 (26) although its popular application to immunoassay dates to Catt and Tregear
The adsorption of proteins on synthetic surfaces, such as polystyrene latex and glass, was studied as early as 1956 (26) although its popular application to immunoassay dates to Catt and Tregear
The adsorption of proteins on synthetic surfaces, such as polystyrene latex and glass, was studied as early as 1956 (26) although its popular application to immunoassay dates to Catt and Tregear
Most probably, The EGn-terminated SAMs are not unique in their ability to resist the adsorption of proteins. SAMs presenting tripropylene sulfoxide groups also prevent the non-specific adsorption of proteins [40].
… Read more
And The adsorption of proteins on synthetic surfaces, such as polystyrene latex and glass, was studied as early as 1956 (26) although its popular application to immunoassay dates to Catt and Tregear
The characteristics of passive adsorption on polystyrene at alkaline pH. (A) Adsorption behavior of seven different proteins in a log–log saturation plot. (▾) Bovine IgM, 1000 kDa; (Δ) bovine secretory IgA, 420 kDa; (◯) ovalbumin, 44 kDa; (□) bovine lgG1, 158 kDa; (□) bovine lgG2a, 152 kDa; (ċ) bovine serum albumin, 69 kDa; (▪) bovine α-lactalbumin, 14 kDa. (B) Adsorptive behavior of the same proteins expressed in a molar percentage bound plot. Dashed lines are extrapolations in the region of constant percentage binding for which empirical data were not reported
Adsorption behavior of seven different proteins in a log–log saturation plot. (▾) Bovine IgM, 1000 kDa; (Δ) bovine secretory IgA, 420 kDa; (◯) ovalbumin, 44 kDa; (□) bovine lgG1, 158 kDa; (□) bovine lgG2a, 152 kDa; (ċ) bovine serum albumin, 69 kDa; (▪) bovine α-lactalbumin, 14 kDa. (B) Adsorptive behavior of the same proteins expressed in a molar percentage bound plot. Dashed lines are extrapolations in the region of constant percentage binding for which empirical data were not reported
Thanks a lot for this energetic participation Dear Dr keshav Bhambri.....
Great suggestion...
Adsorption behavior of seven different proteins in a log–log saturation plot. (▾) Bovine IgM, 1000 kDa; (Δ) bovine secretory IgA, 420 kDa; (◯) ovalbumin, 44 kDa; (□) bovine lgG1, 158 kDa; (□) bovine lgG2a, 152 kDa; (ċ) bovine serum albumin, 69 kDa; (▪) bovine α-lactalbumin, 14 kDa. (B) Adsorptive behavior of the same proteins expressed in a molar percentage bound plot. Dashed lines are extrapolations in the region of constant percentage binding for which empirical data were not reported
As i explored, studies performed at alkaline pH and above the pI of all the proteins studied. Adsorption at alkaline pH was the procedure initially described by Engvall and Perlmann
Whereas, Adsorption at other pHs on polystyrene can give much different patterns. Figure 9.5 illustrates that a linear binding region, i.e., a region in which the proportion of adsorbed IgG (or protein in question) remains constant,
Because the proportion which is adsorbed is a function of the Keq (avidity) of the interaction (3), the increase in the proportion of IgG adsorbed at acid and neutral pH with increasing amounts of added IgG suggests cooperative binding as may occur during aggregation
But the proportion of IgG adsorbed at acid and neutral pH with increasing amounts of added IgG suggests cooperative binding as may occur during aggregation
Right that he increase in the proportion of IgG adsorbed at acid and neutral pH with increasing amounts of added IgG suggests cooperative binding as may occur during aggregation
α-lactalbumin, 14 kDa. (B) Adsorptive behavior of the same proteins expressed in a molar percentage bound plot. Dashed lines are extrapolations in the region of constant percentage binding for which empirical data were not reported
https://www.researchgate.net/deref/https%3A%2F%2Fwww.sciencedirect.com%2Ftopics%2Fchemistry%2Fadsorption-of-protein
https://www.researchgate.net/deref/https%3A%2F%2Fwww.sciencedirect.com%2Ftopics%2Fchemistry%2Fadsorption-of-protein
https://www.researchgate.net/deref/https%3A%2F%2Fwww.sciencedirect.com%2Ftopics%2Fchemistry%2Fadsorption-of-protein
https://www.researchgate.net/deref/https%3A%2F%2Fwww.sciencedirect.com%2Ftopics%2Fchemistry%2Fadsorption-of-protein
Because the proportion which is adsorbed is a function of the Keq (avidity) of the interaction
Adsorptive behavior of the same proteins expressed in a molar percentage bound plot. Dashed lines are extrapolations in the region of constant percentage binding for which empirical data were not reported
