Screening the recognition properties of peptide hormone sequence mutants by analytical high performance liquid affinity chromatography on immobilized neurophysin

1988 ◽  
Vol 53 (11) ◽  
pp. 2627-2636 ◽  
Author(s):  
Giorgio Fassina ◽  
Michal Lebl ◽  
Irwin M. Chaiken
Keyword(s):  
Affinity Chromatography ◽  
High Performance ◽  
Peptide Hormone ◽  
Protein Recognition ◽  
Affinity Matrix ◽  
Structure Modification ◽  
Binding Properties ◽  
Chromatography Method ◽  
Contact Residues ◽  
Critical Contact

Analytical high performance liquid affinity chromatography with immobilized neurophysin was used as a molecular screen to evaluate the effects of peptide hormone structure modification on protein recognition. Immobilization of neurophysin on silica and highly cross-linked agarose occurred with retention of oxytocin and vasopressin binding properties. The effects of one-residue-at-a-time mutation, multi-site sequence simplification, and sequence randomization of critical contact residues were evaluated by extent of binding of the peptides on the affinity matrix. The analytical chromatography method also was used as a stereoselective detector to identify racemic contaminants in peptide hormone preparations.

Standardization of Glycohemoglobin Determinations in the Clinical Laboratory: Three Years of Experience

1992 ◽  
Vol 38 (12) ◽  
pp. 2414-2418 ◽  
Author(s):  
G S Bodor ◽  
R R Little ◽  
N Garrett ◽  
W Brown ◽  
D E Goldstein ◽  
...  
Keyword(s):  
Affinity Chromatography ◽  
Analytical Methods ◽  
High Performance ◽  
Clinical Laboratory ◽  
Hplc Method ◽  
Diabetic Patients ◽  
Years Of Experience ◽  
Chromatography Method ◽  
Before And After

Abstract Measurement of glycohemoglobin has been recommended for the long-term assessment of glycemic control in diabetic patients. Because different analytical methods measure different glycohemoglobin species, it has been difficult to compare results between laboratories. Here we report 3 years of experience with calibration of an affinity chromatography method for measuring total glycohemoglobin (GHb). Calibration was achieved by including in each assay three hemolysate calibrators for which values for HbA1c and GHb had been determined by repeated analyses by high-performance liquid chromatography (HPLC) and affinity chromatography, respectively. Calibration improved interassay precision (CV = 3.20-7.90% and < 5.0% before and after the introduction of calibration, respectively) and eliminated lot-to-lot variability. In 91 samples, HbA1c was estimated by the calibrated affinity chromatography assay and measured by an ion-exchange HPLC method. Estimated and HPLC-measured HbA1c showed no clinically significant differences during 36 months. The high degree of long-term precision, the disappearance of lot-to-lot variability, and the excellent comparability between analytical methods measuring different species of glycated hemoglobins demonstrate the advantages of calibration.

A new method for the analysis of plasmin and plasminogen. Application of high-performance affinity chromatography.

1985 ◽  
Vol 16 (1) ◽  
pp. 23-25
Author(s):  
Kenichi KASAI ◽  
Kiyohito SHIMURA ◽  
Naofumi ITO ◽  
Kohji NOGUCHI ◽  
Mutsuyoshi KAZAMA
Keyword(s):  
Affinity Chromatography ◽  
High Performance ◽  
New Method ◽  
High Performance Affinity Chromatography

Development and Validation of A Reversed Phase-High Performance Liquid Chromatography Method for the Quantitative Estimation of Ramipril Drug Content from Formulated Dosage Form

2016 ◽  
Vol 6 (3) ◽  
Author(s):  
Raju Chandra ◽  
Manisha Pant ◽  
Harchan Singh ◽  
Deepak Kumar ◽  
Ashwani Sanghi
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Active Ingredient ◽  
High Performance ◽  
Limit Of Detection ◽  
Quantitative Estimation ◽  
Reversed Phase ◽  
Uv Detector ◽  
Chromatography Method ◽  
Drug Content

A reliable and reproducible reversed-phase high performance liquid chromatography (RP-HPLC) was developed for the quantitative determination of Remipril drug content from marketed bulk tablets. The active ingredient of Remipril separation achieved with C18 column using the methanol water mobile phase in the ratio of 40:60 (v/v). The active ingredient of the drug content quantify with UV detector at 215 nm. The retention time of Remipril is 5.63 min. A good linearity relation (R2=0.999) was obtained between drug concentration and average peak areas. The limit of detection and limit of quantification of the instrument were calculated 0.03 and 0.09 µg/mL, respectively. The accuracy of the method validation was determined 102.72% by recoveries method.

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