Superior Separation, Resolution and Quantitation with V8 Haemoglobin IEF Analysis
High throughput assay
V8 delivers up to four times the speed
of HPLC Haemoglobin testing
Single format kit
Automated peak detection
For the detection and quantitation of all
major normal and variant haemoglobins
Detection and colour labelling for
crystal-clear result reporting
Gold-standard separation
The only Clinical CE system capable
of Hb IEF separation
A
Next-generation Capillary Electrophoresis technology for better results
Haemoglobin C is a ß chain variant, the second most frequent haemoglobin
variant observed in the western hemisphere. Individuals with homozyosity of
haemoglobin C (ßCßC) only produce haemoglobins C, A2 and F and this results
in a mild to moderate haemolytic anaemia with splenomegaly and an increased
incidence of gall stones. Individuals with only one copy of the haemoglobin C
gene (ßßC) are usually asymptomatic. However haemoglobin C in individuals who
are also heterozygous for the haemoglobin S gene (ßSßC) can also cause sickle
cell anaemia.
C
F
A2
D-Punjab
Haemoglobin D
A
A
Normal Haemoglobin constituents
Haemoglobin is the major protein in the red blood cell; it is the transport
protein for oxygen and is essential for life.
Haemoglobin A. This is the normal, predominant haemoglobin that is
produced from late gestation. Haemoglobin A is a tetramer with two alpha
chains and two beta chains (α2β2 ).
Haemoglobin A2 (HbA2 ) is a normal variant of haemoglobin A that
consists of two alpha and two delta chains (α2δ2 ). and is found in small
quantity in normal human blood.
Superior resolution compared to
competitors systems
Haemoglobin C
Helena Biosciences’ Haemoglobin IFE method is a high
quality, robust test offering the superior separation qualities
of Isoelectric Focusing (IEF) with the ability to both separate
and quantify all major normal and abnormal variants in one
convenient assay format.
The V8’s advanced analysis and automation features offer the
clinician unprecedented screening capabilities.
Superior variant detection
A2
Haemoglobin D-Punjab is a ß chain variant and is one of the most frequently
encountered variants in the western hemisphere. Both the homozygous
(ßD-PunjabßD-Punjab) and heterozygous (ßßD-Punjab) forms are not associated
with any severe symptoms when inherited in isolation from any other
Haemoglobin variants. If co-inherited with Haemoglobin S (ßSßD-Punjab),
sickle cell disease occurs. Homozygotes for Hb D-Punjab produce only
haemoglobins D-Punjab, A2 and F. Heterozygotes for haemoglobin D-Punjab
have relatively lower levels (%) of haemoglobin D than haemoglobin A.
A
A2
Bart’s
ß Thalassaemia
A range of disorders which are characterised by the reduced synthesis of the
Beta chains of Haemoglobin.
A ß-Thalassaemic
F
This abnormal haemoglobin seen during foetal development in individuals with
4-gene deletion alpha thalassemia was characterised at St. Bartholomew’s
Hospital in London, hence Haemoglobin Bart’s. The condition Haemoglobin
Bart’s Hydrops fetalis leads to severe anaemia and oedema. The outcome
of the condition is the death of the foetus in utero or very early neonatal
death. Diagnosis of the condition is by demonstrating the total absence
of Haemoglobin A, A2 and F. The alternative variants detected are
Heamoglobin Bart’s(γ4 ) and sometimes Haemoglobin Portland (ζ2γ2 ).
Bart’s
A
A
Haemoglobin E is a ß chain variant. Haemoglobin E can take homozygous or
heterozygous forms. For individuals with haemoglobin E heterozygosity (ßßE),
approximately 30% of the total haemoglobin is Hb-E (trace pictured) and
approximately 90% of these individuals will have microcytosis, a condition in
which the red blood cells are unusually small. Homozygous (ßEßE) individuals
only have haemoglobin E, A2 and F and do not generally display many more
symptoms, however they often display a more marked microcytosis and also
a mild to moderate anaemia.
F
A2
F
ß-Thalassaemia
intermedia, patients
may require episodic
blood transfusions.
ß-Thalassaemia
Major,
severe hypochromic
mycrotic anaemia.
Also causes
splenomegaly.
A2
Haemoglobin F
E
Haemoglobin E
Haemoglobin F (Hb-F) is the main oxygen transport protein in the foetus
during the last 7 months of development in the uterus, and also in the
newborn to approximately 6 months of age. In the majority of adults
Haemoglobin F is 0.7% or less. However, the control of the proportion
of Hb-F containing cells and the rate of Hb-F synthesis is polygenic and
a significant amount of adults have an Hb-F percentage slightly above
this level. Also 15-20% of pregnant women can see a rise in their Hb-F
percentages with levels as high as 5%.
A2
Capillary-based IEF for unique resolving power and analytical performance
A
Haemoglobin S
Hb-S is a ß chain variant. Individuals who are homozygous for the Hb-S (ßSßS) gene
suffer from a condition known as sickle cell anaemia. This condition leads to a change
in shape of red blood cells which form an irregular crescent shape. This leads to a
large reduction of the life span of the red blood cell and also anaemia, which is due to
the reduced affinity for oxygen of Haemoglobin S. The persistence of the haemoglobin
S gene is thought to be due to the fact that individuals with heterozygous haemoglobin
S (ßßS) have some conferred resistance to malarial parasitisation.
Helena Biosciences Europe Queensway South, Team Valley Trading Estate, Gateshead, Tyne and Wear, NE11 0SD, United Kingdom
A2
A
ß-Thalassaemia
Minor (trait), mycrotic
and possible mild
anaemia, no
health issues normal
iron levels.
trait trace
overlaid onto
normal trace
Tel +44 (0)191 482 8440
Fax +44 (0)191 482 8442
Diffuse mixture of proteins
S
Lower pH
(acid)
Higher
pH
(base)
Haemoglobin detected
by absorbance
S
A
F
F
A2
Haemoglobins of differing
isoelectric points are loaded as a
diffuse mixture into the capillary.
Under voltage, a pH gradient is
created in which Haemoglobins
migrate to their individual
isoelectric points.
Direct detection of the haem
group occurs at 415nm, allowing
the reporting of high-resolution
results.
A2
The resulting Platinum 4V
trace displays excellent variant
separation and quantification.
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