The latest publication from the CuriRx team illustrates and improved HPLC method

June 28, 2018



The latest publication from the CuriRx team illustrates an improved HPLC method to ensure better analyses and better separation of peaks in terms of theoretical plate, peak width, height, and symmetry, along with mitigating impurities’ interaction with the column.




Accurate quantification of the intact proteins, antibodies or peptides and their impurities without interaction to silanols of HPLC column.


Hydroxypropyl ß Cyclodextrin (HPCD) is added in the mobile phase at different concentrations. Different commercial SEC-HPLC columns and biologics with a molecular weight ranging from 5.8 kDa to 150kDa were assessed with and without cyclodextrin.


Addition of non-ionic sugars such as Hydroxypropyl ß Cyclodextrin in the mobile phase, resulted improved peak performance such as theoretical plates, peak resolution, peak width, peak height, and improved quantification of aggregates in biologics such as antibodies Humira and Actemra, and peptides such as insulin. There is an increase in peak height, reduced retention time, increased plate and reduced peak width with increasing concentration of cyclodextrin studied.


High ionic strength, basic amino acids such as arginine, organic solvents (with a concentration low enough not to precipitate protein), sodium perchlorate and ion pairing agents in the mobile phase used for separation of peptides, proteins and antibodies to prevent silanol interaction. These commonly used solutions are not always successful, as they not only interact with the biologic, but are sometimes, not compatible. The non-ionic cyclodextrin itself does not cause protein aggregation but prevents the nonspecific binding or interaction of protein itself and thereby allowing for improved resolution, and accurate quantification of aggregates in antibodies, and peptides. The data on the separation in presence of cyclodextrin in the mobile phase showed higher peak resolution, improved peak shape, accurate apparent molecular weight, improved efficiency, and less peak tailing for biological products.


Hydroxypropyl ß Cyclodextrin in the mobile phase, resulted improved SEC-HPLC resolution, and quantitation of aggregates in biologics by preventing the interaction of biologics to silanol of the commercial SEC-HPLC columns.

Friends and Colleagues,

The government has classified the biopharma industry as essential during this trying time. As such, CuriRx is keeping our doors open for business.

We will continue to provide quality contract R&D services in a timely manner and do our part to contribute patient care. At CuriRx, the health and well-being of our employees, clients, and the general public are still our topmost concern and priority. Internally, we are encouraging all staff to adhere to calls for social distancing, asking that they work remotely when possible and maintain a measure of physical distance while working in the lab.

As this pandemic continues to challenge our communities, we have also implemented a number of additional safety measures around the office, including extra hand sanitizer dispensers, “deep cleaning” of all surfaces and doorknobs with heavy duty disinfectants, and encouraging video conferences even internally to discourage the gathering of too many people in one room at a time.

As always, we will continue to monitor the situation globally and adjust our behavior according to federal, state and public health guidelines.

Stay safe and healthy,

CuriRx, Inc.