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How can I remove acetate from peptides?
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+ Solid phase extraction
+ Peptide chemistry
+ Peptide synthesis
+ Cell line
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Mizanur Rahman
How can I remove acetate from peptides?
I have no idea what will happen! However, an acetylated protein is generally inactive and one of the major degradants or related substance. Thus, the protein may be active (preferable), deactivated, or totally degraded.
I have no idea what will happen! However, an acetylated protein is generally inactive and one of the major degradants or related substance. Thus, the protein may be active (preferable), deactivated, or totally degraded.
Hi, can anybody suggest that what would be the effect of acetate salt on hydrophillic peptides? I want to explore peptides for biological activity, my concern is same that if I remove TFA and replace it wil acetate salt, which might affect its activity. What could be done to prevent it?
Hi, can anybody suggest that what would be the effect of acetate salt on hydrophillic peptides? I want to explore peptides for biological activity, my concern is same that if I remove TFA and replace it wil acetate salt, which might affect its activity. What could be done to prevent it?
If the peptides are hydrophobic then i would say its rather easy to remove the acetate salts (if any) by a simple aqueous work-up (solvent extraction) followed by centrifugations and washings. They remaining acetic acid could be taken care of by co-evaporation as suggested. We have performed, in several occasions, such purification for various assays and never faced any problem whatsoever. There is absolutely no issue in resolubilizing them in toluene followed by evaporation (2-3 times) and lyophilization to get rid of the remaining trace of acetic acid. Hope it helps ! Cheerio !
If the peptides are hydrophobic then i would say its rather easy to remove the acetate salts (if any) by a simple aqueous work-up (solvent extraction) followed by centrifugations and washings. They remaining acetic acid could be taken care of by co-evaporation as suggested. We have performed, in several occasions, such purification for various assays and never faced any problem whatsoever. There is absolutely no issue in resolubilizing them in toluene followed by evaporation (2-3 times) and lyophilization to get rid of the remaining trace of acetic acid. Hope it helps ! Cheerio !
Depends on the pH. If it is very acidic you have the problem of Acetylating organic acid groups (and also deactivate the enzyme). If the pH is ~7 nothing will happen.
Depends on the pH. If it is very acidic you have the problem of Acetylating organic acid groups (and also deactivate the enzyme). If the pH is ~7 nothing will happen.
Hi Somnath Mukherjee, Many thanks for your answer. The peptides are insoluble in water and are very hydrophobic. I think some acetic acid remained in the samples when they were dried down. If the samples are now dry would you suggest resolubilising them in toluene and then rotavap?
Hi Somnath Mukherjee, Many thanks for your answer. The peptides are insoluble in water and are very hydrophobic. I think some acetic acid remained in the samples when they were dried down. If the samples are now dry would you suggest resolubilising them in toluene and then rotavap?
You can try size exclusion chromatography/gel permeation chromatography (sephadex LH-20 can be used in polar organic solvents) or as your compounds are insoluble in water, simply wash the solid peptide with water. best regards
You can try size exclusion chromatography/gel permeation chromatography (sephadex LH-20 can be used in polar organic solvents) or as your compounds are insoluble in water, simply wash the solid peptide with water. best regards
If the peptides are hydrophobic then i would say its rather easy to remove the acetate salts (if any) by a simple aqueous work-up (solvent extraction) followed by centrifugations and washings. They remaining acetic acid could be taken care of by co-evaporation as suggested. We have performed, in several occasions, such purification for various assays and never faced any problem whatsoever. There is absolutely no issue in resolubilizing them in toluene followed by evaporation (2-3 times) and lyophilization to get rid of the remaining trace of acetic acid. Hope it helps ! Cheerio !
If the peptides are hydrophobic then i would say its rather easy to remove the acetate salts (if any) by a simple aqueous work-up (solvent extraction) followed by centrifugations and washings. They remaining acetic acid could be taken care of by co-evaporation as suggested. We have performed, in several occasions, such purification for various assays and never faced any problem whatsoever. There is absolutely no issue in resolubilizing them in toluene followed by evaporation (2-3 times) and lyophilization to get rid of the remaining trace of acetic acid. Hope it helps ! Cheerio !
Dear Niamh Murray and Kamal Malhotra Acetic acid or TFA is a counter ion for positively charged groups/residues (e.g. free amino group, lysine, arginine, histidine) and it will be in stoichiometric amount - the molar content of acetic acid or TFA will be equal to the molar content of those positively charged groups/residues in peptide sample. This is because after freeze drying you always obtain the substance with zero average charge. All excessive acetic acid/TFA will sublime. If you want to remove acetic acid/TFA you can dissolve a peptide in a relatively high volume of solution with other counter ion (e.g. chloride) and freeze dry sample. Do it several times (e.g. three times - number of these steps depend on molar content of your peptide). In that way you will substitute one counter ion with the other by "dilution" of the first with the second. Then you will probably have to remove the excess of e.g. NaCl by standard desalting methods.
Dear Niamh Murray and Kamal Malhotra Acetic acid or TFA is a counter ion for positively charged groups/residues (e.g. free amino group, lysine, arginine, histidine) and it will be in stoichiometric amount - the molar content of acetic acid or TFA will be equal to the molar content of those positively charged groups/residues in peptide sample. This is because after freeze drying you always obtain the substance with zero average charge. All excessive acetic acid/TFA will sublime. If you want to remove acetic acid/TFA you can dissolve a peptide in a relatively high volume of solution with other counter ion (e.g. chloride) and freeze dry sample. Do it several times (e.g. three times - number of these steps depend on molar content of your peptide). In that way you will substitute one counter ion with the other by "dilution" of the first with the second. Then you will probably have to remove the excess of e.g. NaCl by standard desalting methods.
I have no idea what will happen! However, an acetylated protein is generally inactive and one of the major degradants or related substance. Thus, the protein may be active (preferable), deactivated, or totally degraded.
I have no idea what will happen! However, an acetylated protein is generally inactive and one of the major degradants or related substance. Thus, the protein may be active (preferable), deactivated, or totally degraded.
More
VOTE
Hi, can anybody suggest that what would be the effect of acetate salt on hydrophillic peptides? I want to explore peptides for biological activity, my concern is same that if I remove TFA and replace it wil acetate salt, which might affect its activity. What could be done to prevent it?
Hi, can anybody suggest that what would be the effect of acetate salt on hydrophillic peptides? I want to explore peptides for biological activity, my concern is same that if I remove TFA and replace it wil acetate salt, which might affect its activity. What could be done to prevent it?
More
VOTE
How is acetic acid solution added during compounding/manufacturing of a peptide? By pouring, spraying, other?
How is acetic acid solution added during compounding/manufacturing of a peptide? By pouring, spraying, other?
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If the peptides are hydrophobic then i would say its rather easy to remove the acetate salts (if any) by a simple aqueous work-up (solvent extraction) followed by centrifugations and washings. They remaining acetic acid could be taken care of by co-evaporation as suggested. We have performed, in several occasions, such purification for various assays and never faced any problem whatsoever.
There is absolutely no issue in resolubilizing them in toluene followed by evaporation (2-3 times) and lyophilization to get rid of the remaining trace of acetic acid.
Hope it helps !
Cheerio !
If the peptides are hydrophobic then i would say its rather easy to remove the acetate salts (if any) by a simple aqueous work-up (solvent extraction) followed by centrifugations and washings. They remaining acetic acid could be taken care of by co-evaporation as suggested. We have performed, in several occasions, such purification for various assays and never faced any problem whatsoever.
There is absolutely no issue in resolubilizing them in toluene followed by evaporation (2-3 times) and lyophilization to get rid of the remaining trace of acetic acid.
Hope it helps !
Cheerio !
More
VOTE
Depends on the pH. If it is very acidic you have the problem of Acetylating organic acid groups (and also deactivate the enzyme). If the pH is ~7 nothing will happen.
Depends on the pH. If it is very acidic you have the problem of Acetylating organic acid groups (and also deactivate the enzyme). If the pH is ~7 nothing will happen.
More
VOTE
Hi Somnath Mukherjee, Many thanks for your answer. The peptides are insoluble in water and are very hydrophobic. I think some acetic acid remained in the samples when they were dried down. If the samples are now dry would you suggest resolubilising them in toluene and then rotavap?
Hi Somnath Mukherjee, Many thanks for your answer. The peptides are insoluble in water and are very hydrophobic. I think some acetic acid remained in the samples when they were dried down. If the samples are now dry would you suggest resolubilising them in toluene and then rotavap?
More
VOTE
You can try size exclusion chromatography/gel permeation chromatography (sephadex LH-20 can be used in polar organic solvents) or as your compounds are insoluble in water, simply wash the solid peptide with water. best regards
You can try size exclusion chromatography/gel permeation chromatography (sephadex LH-20 can be used in polar organic solvents) or as your compounds are insoluble in water, simply wash the solid peptide with water. best regards
More
VOTE
If the peptides are hydrophobic then i would say its rather easy to remove the acetate salts (if any) by a simple aqueous work-up (solvent extraction) followed by centrifugations and washings. They remaining acetic acid could be taken care of by co-evaporation as suggested. We have performed, in several occasions, such purification for various assays and never faced any problem whatsoever.
There is absolutely no issue in resolubilizing them in toluene followed by evaporation (2-3 times) and lyophilization to get rid of the remaining trace of acetic acid.
Hope it helps !
Cheerio !
If the peptides are hydrophobic then i would say its rather easy to remove the acetate salts (if any) by a simple aqueous work-up (solvent extraction) followed by centrifugations and washings. They remaining acetic acid could be taken care of by co-evaporation as suggested. We have performed, in several occasions, such purification for various assays and never faced any problem whatsoever.
There is absolutely no issue in resolubilizing them in toluene followed by evaporation (2-3 times) and lyophilization to get rid of the remaining trace of acetic acid.
Hope it helps !
Cheerio !
More
VOTE
Tomasz Fraczyk: Thank you so much!
Tomasz Fraczyk: Thank you so much!
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Dear Niamh Murray and Kamal Malhotra
Acetic acid or TFA is a counter ion for positively charged groups/residues (e.g. free amino group, lysine, arginine, histidine) and it will be in stoichiometric amount - the molar content of acetic acid or TFA will be equal to the molar content of those positively charged groups/residues in peptide sample. This is because after freeze drying you always obtain the substance with zero average charge. All excessive acetic acid/TFA will sublime. If you want to remove acetic acid/TFA you can dissolve a peptide in a relatively high volume of solution with other counter ion (e.g. chloride) and freeze dry sample. Do it several times (e.g. three times - number of these steps depend on molar content of your peptide). In that way you will substitute one counter ion with the other by "dilution" of the first with the second. Then you will probably have to remove the excess of e.g. NaCl by standard desalting methods.
Dear Niamh Murray and Kamal Malhotra
Acetic acid or TFA is a counter ion for positively charged groups/residues (e.g. free amino group, lysine, arginine, histidine) and it will be in stoichiometric amount - the molar content of acetic acid or TFA will be equal to the molar content of those positively charged groups/residues in peptide sample. This is because after freeze drying you always obtain the substance with zero average charge. All excessive acetic acid/TFA will sublime. If you want to remove acetic acid/TFA you can dissolve a peptide in a relatively high volume of solution with other counter ion (e.g. chloride) and freeze dry sample. Do it several times (e.g. three times - number of these steps depend on molar content of your peptide). In that way you will substitute one counter ion with the other by "dilution" of the first with the second. Then you will probably have to remove the excess of e.g. NaCl by standard desalting methods.
More
VOTE
Peptide contains basic residues (charge 8)
Peptide contains basic residues (charge 8)
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