Added application data No.114
「Analysis of intact proteins」
『Analysis of peptides/proteins and monoclonal antibodies by UHPLC』
This technical report focuses on the analysis of proteins and monoclonal antibodies. Moreover, it contains information from intact to fragmented peptides.
In addition to C1, C8, C18 were added to the lineup.
Here are the results that can only be obtained with 300Å.
Column:
Develosil UHPLC C30, 1.6um (2.0x50mm)
Column:
Develosil UHPLC C30, 1.6um (2.0x50mm)
We analyzed peptides using “Develosil UHPLC series” announced on Tuesday, October 23, 2018.
A major feature is that it is possible to change the analytical conditions that used ion-pairing reagents such as TFA until now to mild mobile phase such as ammonium formate.
Changing the pH from 3 to 7 makes it possible to correspond to LC / MS negative mode as well.
Column:
Develosil HSR C18 Peptide, 5um (4.6x150mm)
Develosil HSR C18 Peptide, 3um (4.6x75mm)
Amyloid β-protein is a peptide of about 40 amino acids, and it is said that accumulation of this substance is closely related to Alzheimer’s disease.
We analyzed amyloid β-protein using a Corona detector this time. In addition, this method can also be used for LC / MS by changing the column inner diameter to 2.0 mm.
It is an analysis example in which insulin, BSA, monoclonal antibody, thyroglobulin are digested using a convenient kit.
In order to obtain more information, 2um particle size may be used, but in this data it is an analysis example using 3um particle size and a general-purpose HPLC system, so it is easy to analyze digests.
This is an example of analysis of oligodeoxythymidine using HPLC and UHPLC system.
Simply by selecting the column size and particle size according to your system you can easily analyze oligonucleic acids.
This data is not data transferred method from HPLC to UHPLC.
・Sample
dCTP, dTTP, dGTP, dATP
・Column
Develosil HSR AQ C18, 5um (4.6x150mm)
Develosil HSR AQ C18, 2um (2.0x50mm)