Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of... – Animal cell – per se – expressing immunoglobulin – antibody – or...
Patent
1996-03-27
2000-05-16
Hutzell, Paula K.
Chemistry: molecular biology and microbiology
Animal cell, per se ; composition thereof; process of...
Animal cell, per se, expressing immunoglobulin, antibody, or...
435331, 435334, 435346, 5303882, 530399, 5303911, 5303913, C12N 1585, C12N 512, C07K 1600
Patent
active
060636203
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to a novel hybridoma cell line, in particular it relates to a novel hybridoma cell line which secretes monoclonal antibodies capable of binding to the AT.sub.1 subtype of the Angiotensin II receptor (AT.sub.1 receptor). The invention also relates to monoclonal antibodies secreted by the hybridoma, which antibodies may be used in a test kit having several diagnostic and monitoring applications. It also relates to the use of the monoclonal antibodies in therapeutic applications such as the control of smooth muscle stimulation.
The hormone angiotensin II (Ang II) forms part of the renin--angiotensin system which helps to control electrolyte balance and blood pressure within the body. There are several tissues within the body upon which Ang II acts, they include the adrenal gland, uterus, liver, brain and kidney.
Amongst the several established functions of angiotensin II, it is known to stimulate smooth (unstriated) muscle cell contraction. It stimulates the contraction of smooth muscle cells in the blood vessel wall thus causing vaso-constriction, which leads to hypertension. Most treatments for high blood pressure will include blockage of angiotensin function in one way or another. Smooth muscle also occurs in other locations, for example in the uterus and in the gastrointestinal tract, and elsewhere.
Ang II also stimulates the secretion of aldosterone by the adrenal cortex. Aldosterone is a potent hormone which acts primarily on the kidney to promote sodium retention and thus inter alia, heightens the hypertensive effects of angiotensin acting directly on the vasculature.
Ang II is known to act on various sites in the brain, and one of its actions in animals is the regulation of thirst and drinking.
Angiotensin also has trophic effects on the vasculature, promoting growth of the muscles in the arterial wall. It is also thought to be angiogenic, i.e. it causes vascularisation of newly developing tissue.
The actions of angiotensin II in cells are mediated through two important intracellular signalling mechanisms. When the hormone binds to its receptor, it activates a specific enzyme, phospholipase C, which acts upon a constituent of the cell membrane called phosphatidyl inositol. This is split by the enzyme into two moieties, called inositol trisphosphate (IP3), and diacylglycerol (DAG). Both of these are involved in eliciting further effects within the target cell. IP3 stimulates increased cellular cytosolic calcium concentrations, which in turn evokes other cellular responses, whereas DAG stimulates another specific enzyme called protein kinase C (PKC).
Most of the established effects of Ang II have been found to occur via the AT.sub.1 subtype of the Ang II receptor, which is a seven transmembrane domain receptor. This receptor has been cloned and sequenced from a variety of tissues, and has been found to be a 359 amino acid polypeptide with a predicted molecular weight of around 40 kD (Bernstein and Alexander, (1992), Endocr. Rev., 13, 381-386). Studies using photo-affinity labelling and crosslinking agents have suggested molecular weights for mature receptor of approximately 65 kD and 116 kD, respectively, which may reflect glycosylation of asparagine residues within the extra-cellular domain.
Whilst polyclonal antibodies and anti-idiotypic antibodies have been prepared to the Ang II receptor, from which it has been postulated that the receptor has a molecular weight of from 60 to 95 kD, and 63 kD respectively, to date no-one has succeeded in preparing a monoclonal antibody to this receptor.
Garcia et al in Science, 257, 502-507, (1992) described the preparation of a monoclonal antibody to the hormone Ang II itself.
Couraud in J. Immunol., 138 (4), 1164-8, (1987) described the preparation of anti-idiotypic serum from a rabbit immunized with one anti-angiotensin II monoclonal antibody. Pfister et al in Regul. Pept., 44 (2), 109-17, (1993) described the preparation of anti-idiotypic antibody that bound to the angiotensin II receptor. The preparation of polyclonal antibodies to
REFERENCES:
Eshhar et al (1985) Monoclonal Antibody Strategy and Techniques in Hybridoma Technology in the Biosciences & Medicine. Ed Timothy Springer pp. 3-25.
Zelezna et al Biochemical and Biophys. Research Comm. vol. 183, No. 2 (1992) pp. 781-788. (Submitted by Applicant).
Barker Stewart
Vinson Gavin Paul
Bansal Geetha P.
Hutzell Paula K.
Queen Mary & Westfield College
LandOfFree
Type I angiotensin II receptor specific monoclonal antibodies an does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Type I angiotensin II receptor specific monoclonal antibodies an, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Type I angiotensin II receptor specific monoclonal antibodies an will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-258019