New Cytokines CE ELISA kits
Launching File
Part4: Cell Surface Antigens
Products:
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cat#
Product Description
KAPMS108
human sp185HER-2
KAPMS109
human sCD-44
KAPMS110
human sIL-2R
KAPMS112
human sCD-23
KAPMS115
human sCD30
ELISA
96 Tests
Table of contents:
1. Overview
2. Cell surface antigens products
3. Technical Performances
4. Package Inserts
Cytokines CE- Part4- Cell Surface Antigens
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1. Overview
Both T and B cells have surface antigens that are characteristic of different stages in their life
cycle, and antibodies have been prepared that identify the antigens. Knowledge of the specific
type and stage of maturation of the tumour cells helps physicians determine the prognosis
and course of treatment for the patient.
2. Cell Surface Antigens- Clinical background
1. sp185HER-2
ELISA KIT
Introduction
The human HER-2 (c-erbB-2, new) gene encodes a putative transmembrane growth factor receptor (p185
protein) that is closely related to the epidermal growth factor receptor protein. The HER-2 gene product is a 185
kDa-glycoprotein that contains an extracellular ligand-binding domain and intracellular tyrosine kinase
activity (9). p185HER-2 protein staining is observed only in low levels in epithelial cells of most organs in normal
human tissues and at slightly higher levels in fetal tissues.
Both HER-2 oncogene amplification and oncoprotein overexpression have been analyzed for potential utility in
diagnostic and prognostic tests for: breast (5, 6, 8, 11, 14, 18, 19, 21, 23), ovarian (3, 15, 17, 19, 26), gastric (10, 25),
lung (7, 12, 22); and other cancers (1, 4, 5, 16, 20, 24). In these malignancies the p185 HER-2 oncoprotein
overexpression is correlated with a poor prognosis.
In 15-40 % of primary breast cancers, amplification of the HER-2 oncogen is found which is highly correlated
with overexpression of the encoded 185 kDa protein and seems to play a major role especially during the
initiation of ductal carcinomas. p185 HER-2 overexpression is described as independent prognostic factor with
greater predictive power than most of the currently used prognostic tools (21) - especially in axillary lymphnode-positive breast cancer patients.
Clinical applications
Studies analyzing small series of patients have suggested a prognostic value for p185 HER-2 oncoprotein
expression in axillary node negative (ANN) patients. An association between oncoprotein expression and
decreased overall survival among ANN patients with good nuclear grade tumors has been demonstrated (13).
In addition it has been reported that in low risk patients (estrogen receptor positive, small tumors), p185HER-2
expression was associated with early recurrence (2). Data demonstrate the large body of evidence implicating
p185HER-2 oncoprotein in the biology and prognosis of breast carcinoma.
32 % of ovarian carcinomas overexpress the p185HER-2 oncoprotein. Survival of those patients is significantly
worse compared with cases of normal p185 HER-2 protein expression. Additionally, patients whose tumors have
high p185 protein expression are significantly less likely to have a complete response to primary therapy. Also
non-small cell lung cancers which express the p185HER-2 protein do so at higher levels than those found in
normal bronchial epithelium, and expression in adenocarcinoma of the lung is independently associated with
diminished survival (7). A correlation between p185 HER-2 expression, and clinical outcome has been also
demonstrated for head and neck, salivary gland and placental carcinomas.
p185HER-2 is useful in identifying cancer cells with increased aggressiveness. Soluble p185HER-2 protein levels in
serum can be used as diagnostic tool for monitoring the extent of tumor spread, postoperative relapse and/or
metastatic risk for different cancers.
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2. s CD-44 ELISA KIT
Introduction
CD44 (Pgp-1; Ly-24; ECMR III; F10-44-2; H-CAM; HUTCH-I; In(Lu)-related p80; Hermes antigen; hyaluronan receptor) is a
polymorphic glycoprotein with apparent molecular weights ranging from 85 kDa to 250 kDa. This cell membrane
associated molecule has a cytoplasmic tail (mediates the interaction with the cytoskeleton), a short hydrophobic
transmembrane region and an NH2-terminal extracellular (binds to hyaluronate) domain.
CD44 isoforms participate in a wide variety of cell-cell or cell-matrix interactions including lymphocyte homing,
establishment of B and T cell immune responses, tumor metastases formation and inflammation.
Three isoform categories of the CD44 molecule have been identified:
1) an 80-90 kDa isoform, the so-called calibrator form named CD44std, which is widely distributed on several
hematopoietic and nonhemato-poietic cells including all subsets of leukocytes, monocytes, erythrocytes, many types of
epithelium, mesenchymal elements like fibroblasts, smooth muscle cells and glial cells of the central nervous system,
2) a medium size category of 110-160 kDa which is weakly expressed on epithelial cells and highly expressed in some
carcinomas and
3) a category which includes very large isoforms of 250 kDa covalently modified by the addition of chondroitin sulphate
(12).
These bigger isoforms of CD44 arise by alternative splicing of one or more "variant" exons (v2-v10) into the extracellular
part of the 90kDa constant form molecule. Compared to the calibrator CD44, all larger isoforms are expressed in a much
more restricted fashion, only in a few normal tissues or on the surface of certain tumor cells. Some splice variants of CD44
play important and distinct roles in tumor metastasis .
The sCD44std ELISA detects all circulating CD44 isoforms comprising the calibrator protein sequences (black area).
cytoplasmic
extracellular
transmembrane
N
V2
CD44 protein:
V3
V4
V5
V6
V7
V8
C
V9 V10
- standard protein sequences (black area)
- variant exons (open boxes numbered v2 - v10)
Clinical applications
-
-
-
-
-
colorectal carcinomas: in human colorectal neoplasia CD44 variant proteins are found on all invasive
carcinomas and during carcinoma metastasis. Variants are already expressed at a relatively early stage of
colorectal carcinogenesis and tumor progression.
gastric cancer: tumors from patients suffering from stomach adenocarcinomas express CD44 variants.
Adenocarcinomas of the intestinal type are strongly positive for exon v5 and v6, whereas diffuse type
adenocarcinomas predominantly express exon v5.
lung, breast cancer: in malignant tissues there is gross overproduction of alternatively-spliced large
molecular variants in all samples, whereas in the control samples only the calibrator product was routinely
detected with occasional minimal quantities of one or two small variants.
lymphoma: in gastrointestinal lymphoma overexpression of CD44 has been correlated with poor survival
and more disseminated disease .
Overexpression of CD44 is also found in several aggressive, but not low-grade, non-Hodgkin´s lymphomas
(7) as well as in Hodgkin´s and nodal diffuse lymphomas .
tonsil, skin cancer: variant CD44 isoform expression can be demonstrated in the plasma membrane of
squamous cells of skin and tonsil epithelial and is greatly diminished in malignant squamous epithelial
tumors .
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HIV: CD44 is almost completely depleted from the surface of HIV-infected cells .
inflammatory joint diseases: CD44 expression was decreased in synovial fluid neutrophils from most
patients .
3. sCD-25 (IL-2R) ELISA KIT
Introduction
Interleukin-2 (IL-2) has been described as a factor that promotes the growth and proliferation of human T-cells thus
occupying a pivotal role in the generation of the immune response. This proliferation of T lymphocytes is triggered by the
interaction of IL-2 with its specific cell surface receptor following T lymphocyte activation. The receptor for IL-2 is
composed of at least three distinct polypeptide subunits , the IL-2R a, IL-2R b , and the IL-2R  chains giving rise to a 55-65
kDa membrane bound protein.
The genes encoding IL-2 and these three receptor subunits have been cloned and their complete primary structures have
been deduced . Evidence has accumulated that suggests a critical role of IL-2R  in IL-2 signal transduction (28). IL-2R
coupled tyrosine kinases are believed to play a crucial role in that signal transduction.
In addition to the de-novo expression of IL-2R by activated peripheral blood T-cells, a released and fully soluble form of IL2R (sIL-2R) has been detected . It was shown that sIL-2R is present in vivo, at low levels in the sera of healthy persons and at
markedly elevated levels in various pathological conditions like neoplastic disorders. The structural, functional and
molecular characteristics of sIL-2R have been extensively investigated. By virtue of its ability to bind IL-2, this soluble
molecule plays a role in the regulation of the immune response. The detection and quantitation of sIL-2R provides clinicians
with a useful and simple means of assessing immune function in vivo as part of the investigation, management and
prognosis of a broad spectrum of human diseases.
Clinical applications
- hematologic malignancies (neoplasias):
Soluble IL-2R levels are significantly elevated in the sera of patients with adult T-cell leukemia, hairy cell
leukemia , lymphocytic leukemia , Hodgkin's and Non-Hodgkin's lymphoma , liver, breast and lung cancer
.- autoimmune or inflammatory diseases:
In patients with systemic lupus erythematosus, rheumatoid arthritis , progressive systemic sclerosis ,
polymyositis , Kawasaki disease , multiple sclerosis and Diabetes type I plasma concentrations of sIL-2
receptor are significantly elevated as compared to controls.
- infectious diseases:
Both viral and mycobacterial infections lead to high levels of soluble IL-2R found in Hepatitis , HIV-1 ,
infectious mononucleosis, measles , leprosy, tuberculosis as well as in malaria .
- transplantation or rejection:
sIL-2R turned out to be a very useful marker for monitoring of transplant recipients who show significant
elevations of sIL-2R serum levels experiencing rejection episodes in kidney , lung , heart and liver
transplantations.
- chronic renal failure or dialysis:
Impaired renal function elevates serum sIL-2R as this molecule is actively transported into the urine .
- skin:
sIL-2R is significantly elevated in patients with thermal injury .
- therapy:
Recombinant IL-2 as a cancer therapeutic can provoke release of sIL-2R.
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4. sCD-23 ELISA KIT
Introduction
CD23 is described as a 45 kDa protein found on the surface of IgM bearing B cells, eosinophils, macrophages and some T
and NK cells. It is also found on EBV-transformed B cells. Additionally, a released form has been described . When first
released the CD23 molecule is 35 kDa; however, this form is quickly cleaved to obtain the more stable, soluble form which
is 25 kDa in size (3). Recently the structure of the CD23 molecule was characterized by cloning and sequencing techniques .
Soluble CD23 has been shown to be the B cell growth factor (BCGF). Soluble CD23 is also referred to as Blast-2 and as the
low affinity IgE receptor (FCeRII) (1). It has been speculated that CD23 may up-regulate IgE synthesis in conjunction with T
cell promoted interleukin-4 ; however, the specific physiologic role of this molecule is not yet well understood.
Clinical applications
Elevated levels of CD23 have been found in research studies of samples from people with B cell-derived Chronic
Lymphocytic Leukemia (B-CLL), with Hyper IgE Syndrome and post-Bone Marrow Transplantation samples . CD23 levels
may be proven to relate to disease course in Hairy Cell Leukemia (HCL).
5. sCD-30 ELISA KIT
Introduction
The CD30 (Ki-1) molecule was identified by a monoclonal antibody which was originally found to react with an epitope
present in Hodgkin's and Reed-Sternberg cells in Hodgkin's disease. Later, the Ki-1 antigen was found to be consistently
expressed by a subgroup of diffuse large-cell lymphomas that were called Ki-1 positive (Ki-1+) anaplastic large-cell
lymphomas (ALCL) .
Characterization of the CD30 antigen has shown it to be in its mature form a transmembrane protein of about 120kDa (12,
22) elaborated from an 84kD cytoplasmic precursor primarily through glycosylation . The cloning of the CD30 gene has
allowed the identification of a cDNA with an open reading frame predicting a 595 amino acid polypeptide. The
extracellular domain of CD30, comprising 365 residues, has proved to be homologous to that of the TNF-receptor
superfamily . The CD30 gene is localized at chromosome 1q36 (11), closely linked to other members of the TNF receptor
superfamily comprising TNF-receptors, nerve growth factor, CD40, APO-1/Fas, CD27, OX40 and the neurotrophin receptor.
The CD30 ligand (CD30L) has been identified, showing significant homology to TNF, TNF, FasL, CD40L, CD27L and 41BBl . CD30L is expressed on activated T-cells. Interactions of the cytokine receptor CD30 with its ligand induces
pleiotropic biologic effects, such as differentiation, activation, proliferation and cell death . In CD30 + ALCL cell lines
binding of CD30L induces apoptotic cell death . CD30 furthermore seems to be involved in the control of the CD40/CD40L
signal, T-cell proliferation and B-cell maturation induced by T-cell cytokines. Thus, CD30 seems to transmit information
that is essential for the immune response.
CD30 expression is strictly dependent on activation and proliferation of T- and B-cells. In pathological conditions, CD30
positivity is regarded as a peculiar attribute of Hodgkin's and Reed-Sternberg cells.
There is growing evidence for a potential role of the CD30 molecule in clinical use and therapy . An 85kDa soluble form of
the CD30 molecule (sCD30) has been shown to be released by CD30+ cell in vitro and in vivo .
It is probably derived from the 120kDa membrane bound molecule by proteolytic cleavage . Serum sCD30 detection can be
regarded as a marker of the amount of CD30 + cells present in the body.
Clinical applications
Increased serum levels of sCD30 have been reported for patients with CD30 + ALCL and CD30+ embryonal carcinoma of
the testis and were found to correlate with the clinical phase of the disease, i.e. presentation complete remission (CR),
relapse. Elevated serum values of sCD30 are shown in the majority of patients with Hodgkin's Disease which again
correlate with the presence of B symptoms and with the stage of the disease, i.e. tumor burden.
While elevations of the soluble CD30 in serum of patients affected by infections diseases usually are not detected, infectious
mononucleosis is a notable exception . Serum levels of sCD30 are also increased in most patients with HBsAg-positive
chronic hepatitis and signs of active HBV replication, thus there is association of the raised sCD30 levels with the active
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Cardiov
ascular
& Salt
Balance phase
of the illness. Abnormal soluble CD30 serum accumulation has been reported in Omenn's syndrome, a severe
immunodeficiency . High elevations of sCD30 levels are found in patients of systemic lupus erythematosus which correlate
with disease activity , in patients with the autoimmune liver disease primary biliary cirrhosis and in patients with
rheumatoid arthritis .
3. Cell Surface Antigens Products-Technical performances
human sp185HER-2
ELISA
Cat. Nr.
Label Format
sCD44std ELISA
Controls
Incubation Time
Sensitivity
sCD30 ELISA
KAPMS112
KAPMS115
KAPMS110
HRP-conjugate
HRP-conjugate
HRP-conjugate
0,16 - 10 ng/ml
0,12 - 4 ng/ml
0,31 - 20 ng/ml
12,5 - 400 U/ml
1,6 - 100 ng/ml
10 µl (to be diluted 1:60)
50 µl
100 µl
25 µl
Serum, Plasma
Serum, Plasma
Serum, Plasma
Serum, Plasma
Serum, Plasma
2 vials, lyophilized
no
no
2 vials, lyophilized
no
195 min with
195 min with
195 min with
Biotin-SAV
HRP-conjugate
conjugate
Concentrated (to be diluted)
1/20)
Sample Type
sCD23 ELISA
KAPMS109
15 µl (to be diluted
Sample Size
(CD-25)
KAPMS108
Standard Format
Standard Range
sIL-2R ELISA
135 min with shaking
at 100 rpm
195 min with shaking
shaking at 100 rpm shaking at 100 rpm shaking at 100 rpm
0.06 ng/ml
0.015 ng/ml
0.27 ng/ml
6.8 U/ml
0.33 ng/ml
Intra-assay CV
1.9%
4.8%
7.0%
4.0%
4.1%
Inter-assay CV
5.8%
4.1%
12%
6.3%
5.6%
4. Package inserts
The package inserts will be available on BioSource website.
For more information on those products, please visit our website:
www.biosource-diagnostics.com
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