HEMOGLOBINOPATHIE

S ON IEF AND HPLC

Dr. Ahlam Nasser, MD, MMED
Globin switch during in
utero and post-natal life
Normal and Variant Hemoglobin at Birth
and in Older Children
Methods for Hemoglobinopathy screening
and diagnosis
1. Isoelectric focusing
2. HPLC
3. Cellulose Acetate Electrophoresis (Alkaline)
4. Citrate Agar Electrophoresis (Acid)
5. Alkaline Globin Chain Electrophoresis
6. Capillary Zone Electrophoresis
7. Molecular methods
1. Restriction Fragment Length Polymorphism (RFLP),
2. Allelic Discrimination using Real Time PCR end point data, and
3. DNA Sequencing. DNA
Concept of Protein Electrophoresis
Example of IEF 1
High-performance Liquid
Chromatography
Hb SS
Sickle trait
Beta Thalassemia trait
Hemoglobin S with Beta thalassemia
Beta Thalassemia Major
Hb E trait
Hb E disease
References
• https://www.mun.ca/biology/scarr/Hemoglobin_Electrophoresis.html
• Patterns of hemoglobinopathies diagnosed by high-performance
liquid chromatography in and around Pune (Western Maharashtra,
India): A pilot study – by Buch Archana et al.
• Hemoglobinopathies: Current Practices for Screening, Confirmation
and Follow-up - CDC