Proteins expressed in the membrane of B cells represent compelling targets for therapeutic monoclonal antibody development. Although drugs are commercially available, for example, Rituxan®, which targets CD20, a cell surface protein present in all B cells, there is currently no therapy that selectively acts on activated B lymphocytes. (more...)

Proteins have found utility for numerous commercial and clinical purposes, including use in biochemical and chemical processes, and as agents for the treatment and prevention of human and veterinary disease. A major challenge associated with the use of proteins is their inherent instability. Many proteins rapidly degrade in response to "environmental stresses," such as changes in temperature, pH, light, and desiccation, which has implications for their production, transport, use and storage. Attachment of poly(ethylene glycol) to therapeutic proteins, a process commonly referred to as PEGylation, has been used successfully to increase their stability in vivo by reducing both protease degradation and renal clearance. However, PEGylation does not necessarily increase protein stability in response to environmental stresses. The development of a technology that enhances the stability of proteins to such stresses would dramatically increase the number of proteins that could be used commercially, reduce costs associated with protein production, storage and transportation, and increase protein shelf-life. (more...)

The Human Immunodeficiency Virus (HIV) has evolved a number of mechanisms of evading the human immune system.  One way is through a high level of mutation, which makes it difficult to develop a vaccine that stimulates protective immunity against all of the different HIV variants.  Therefore, scientists are searching for a general surrogate marker that could be used to target any HIV-infected cell regardless of its mutational status, enabling eradication of the virus. In this regard, scientists at UCSF have recently begun to take a closer look at Human Endogenous Retroviruses (HERVs) that are present in all human cells.  HERVs are a family of retroviruses found in the human genome and are thought to have originated from an ancient retrovirus that become permanently integrated with the DNA of its host's germ cells.  HERV viruses are inactive in normal cells but one type of HERV, HERV-K, is activated in HIV-infected cells.  The HERV-K proteins are presented on the surface of HIV-infected cells.  Because HERV-K is expressed in all HIV-infected cells, it is thought HERV-K antigens presented on the surface could be a good candidate to generally target any HIV+ cell. (more...)

The kinesin superfamily is an extended family of related microtubule motor proteins, encompasssing a number of families that exhibit a variety of microtubule motor functions, e.g., vesicle and organelle transport, mitotic spindle function, and meiotic spindle function. These proteins typically work as monomers, are ATP dependent, and have plus end-directed microtubule motor activity involved in fast anterograde organelle transport in neurons. (more...)

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Isolation of purified T-cell subsets through typical separation techniques is difficult when dealing with small numbers of cells. A solution is to add bi-specific antibodies that selectively expand a desired subset. For example, to obtain purified CD8+ T-cells, a CD3 and CD4 bi-specific antibody added to a mixed population will act by triggering cell death in CD8+ T-cells and activating cell division in CD4+ T-cells, thus selectively expanding the CD4+ T-cell population. A million PBMC typically can yield 15 to 20 million pure CD4+ or CD8+ T-cells by this approach, which is fully functional for immune response measurements and other experiments. For studies dealing with limited cells that preclude separation, this technique can provide large numbers of polyclonally expanded T-cells. This approach has been applied in several published studies, such as one examining HIV-specific CD8+ T-cells from rectal biopsies, where very few cells are obtainable (Ibarrondo et al, Journal of Virology 2005, 79:4289-97).The limitation to this approach has been the lack of widely available bi-specific antibodies, which currently are produced by purifying the appropriately paired heavy and light chain antibodies from fused hybridomas (e.g. an anti-CD4 and anti-CD8 hybridoma fusion). This is very labor-intensive and expensive. Thus there is the need for an easier method that can produce bi-specific antibodies on a large scale for broader availability to researchers. (more...)

Researchers at UCLA have developed a polyclonal antibody that specifically recognized histone H3 lysine 18 acetylation. The antibody has been tested and confirmed to work properly in multiple assays including western blotting, immunofluorescence, immunoprecipitation, flow cytometry and chromatin immunoprecipitation (ChIP) in both human cells and yeast. (more...)

A novel DNA-based switch that enables the one-step quantitative detection of antibodies in complex samples (such as whole blood) and effectively reduces analysis time from a few hours to less than 5 minutes. (more...)

Despite the numerous advantages inherent to dynamic bead-based microfluidic arrays, current microparticle trapping methods remain limited. There are currently two fundamental classes of microarrays: static and dynamic microarrays. Static microarrays consist of bio-molecules or chemicals immobilized on a static substrate. Alternatively, dynamic microarrays consist of bio-molecules or chemicals immobilized on mobile substrates, such as microparticle. To enable resettable microfluidic arrays, investigators at University of California at Berkeley have developed a novel reusable dynamic particle-based microarray – termed ‘Microfluidic Ping Pong’ (MPP). In contrast to current dynamic microarray techniques, this system can achieve (i) high-density/throughput microparticle trapping, (ii) microdevice resettability, and (iii) microparticle resettability. High-density trapping enables the acquisition of high numbers of data points (i.e. immobilized microparticle) from a single experiment, without sacrificing device ‘real-estate.’ Dynamic microarrays offer a superior platform due to several advantages compared to static microarrays, including faster reaction times due to larger surface areas of the microparticles, reduced background noise, and the ability to ‘mix-and-match’ particles corresponding to different screenings. Also, the constant mixing of solutions and particulate substrates in microfluidic channels results in faster reaction kinetics compared to the diffusion-based mixing of static systems. (more...)

Hepatitis C virus (HCV) afflicts about 170 million people worldwide and yet the mechanisms responsible for the entry and trafficking of HCV into cells are poorly understood. HCV induces an acute illness and often leads to chronic hepatitis and in some cases may lead to hepatocellular carcinoma and/or cirrhosis. There currently is no treatment that the majority of patients with HCV respond to, and those that are given interferon plus ribaviron often display serious adverse side effects. (more...)

Researchers at UCLA have developed a monoclonal antibody that recognizes the human ABL2 (Abelson murine leukemia viral oncogene homolog 2) protein. It has been successfully used in standard immunoblot and immuno-precipitation techniques. (more...)

The use of antibodies to target tumor cell-associated antigens for diagnostic and therapeutic purposes has been a critical step forward in cancer research. As protein engineering capabilities grow, researchers modify antibodies to alter inherent characteristics, such as affinity and immunogenicity, for enhanced imaging and tumor response. One example of this is in the conjugation of various radionuclides to small recombinant antibody fragments (i.e. diabodies and minibodies) for in vivo tumor cell targeting applications. However, it is not always advantageous to use radioactivity, and thus alternative detection systems are necessary. To that end, the search for high-sensitivity and high-specificity probes that circumvent the limitations of organic dyes and fluorescent proteins has led to the discovery and utilization of quantum dots, nanometer-sized semiconductor particles. Quantum dots are brighter than traditional chromophores, have greater stability, and can be used in multiplex imaging due to size-tunable emission wavelengths. To date, bioconjugates with quantum dots are coupled to intact antibodies whose large size makes it difficult to penetrate tissues and tumors. Therefore, it would be advantageous to monitor tumors with a robust, but small, bioconjugate for tandem in vivo monitoring and treatment. (more...)

G2A, initially an orphan GPCR, was identified in a search for downstream transcriptional targets of the oncogene BCR-ABL tyrosine kinase. G2A belongs to a family of sequence related GPCRs that were initially thought to bind to proinflammatory lipids. However it was recently discovered that G2A and its related GPCRs act as proton sensors that increase the acid-induced production of secondary messengers such as inositol phosphates and cyclic AMP.G2A is expressed mainly in immune cells including T and B-lymphocytes, monocytes, macrophages and dendritic cells. Currently, G2A and its related GPCRs are implicated in a variety of diseases including autoimmune disorders, inflammation and cancer. However the exact biological functions of these GPCRs have not been elucidated. Therefore, readily available research tools such as those generated by the UCLA investigators could significantly accelerate the research to better understand the role of these GPCRs under physiological and pathological conditions. (more...)

Brutons tyrosine kinase (Btk) is a kinase enzyme that plays an important role in B cell stimulation and maturation. B cell maturation and function is impaired when there is a Btk mutation, leading to X-linked agammaglobulinemia (XLA). It is known that B cell stimulation occurs after the activation of Btk, which is correlated with an increase in the phosphorylation of the regulatory Btk tyrosine site, Btk Y223. Currently, the only method for distinguishing the phosphorylation site is through the process of metabolic radiolabeling of phosphates, enzymatic digestion of the Btk protein, and then the separation of the resulting phosphopeptide fragments (phosphopeptide mapping). Phosphopeptide mapping is a difficult and time-consuming technique where only a few samples can be processed. There is a commercially available anti-phosphotyrosine antibody that recognizes the phosphorylated site through immunoblot and immunoprecipitation assays. However, there is not an antibody that has more specificity to the regulatory Btk 223 tyrosine site. There is a need for a more specific and easier technique of distinguishing the Btk tyrosine site, and consequently for the detection of B cell stimulation and maturation. (more...)

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Polyclonal antibodies to STAT1 have been prepared in rabbits and are suitable for use in immunoassays. (more...)

Kinesin proteins are used by neurons for transport along microtubules. The A protein is found throughout neurons while the B protein is enriched in dendrites. Antibodies were raised against these two newly discovered kinesins with amino acid similarity, but different localizations. The antibodies are affinity purified and were raised again fusion proteins containing amino acids 1124-1355 (A) and 1135-1419 (B). (more...)

An antibody to the C terminal fragment of the Golgi protein GRASP65, acting intracellularly, prevents entry of cells into mitosis. Targeting GRASP65 in the Golgi could be a novel way to screen compounds for anti-cancer activity by preventing the fragmentation of the Golgi and thereby preventing entry of cells into mitosis. (more...)

Heart attack and stroke are clinical consequences of atherosclerosis, an inflammatory disease of arteries initiated by lipid accumulation in the artery wall. Current animal models for atherosclerosis primarily use Apolipoprotein E (ApoE) and low density lipoprotein receptor (LDLR) knockout mice, or hyperlipidemic rabbits.. Although these models are useful once lead compounds are being tested, early assessment of new drug candidates is impractical due to the cost, slow throughput, limitations of post mortem analysis of lesions, and poor in-vivo imaging technologies that typically require use of radioactive tracers. An animal model that could provide reasonably high throughput, where the development of atherosclerotic lesions or their regression can be easily monitored while the animal is still alive, would provide significant improvement in the ability to obtain physiological information about early stage candidate compounds. (more...)

Monoclonal Antibodies Against the Prokaryotic Cyclic Nucleotide-Modulated Ion Channel Mlok1 (more...)

Recently, activation of the P2Y12 G-protein coupled receptor (GPCRs) has been shown to be central to platelet aggregation. Drugs preventing platelet aggregation are being tested, but one that would be specific to the P2Y12 receptor would capture a large market share. Developing drugs for the P1Y12 receptor is difficult, because it is a receptor that is naturally activated by ADP. Since practically every cell expresses ADP-activatable receptors, developing a drug screening program directed specifically at the P2Y12 receptor has not been possible. (more...)

Multiple Sclerosis (MS) is a devastating neurological disease. MS patients often experience neurologic dysfunction believed to be the result of infiltration of immune cells, inflammation of the central nervous system and eventual nerve damage. (more...)

BRCA2/RAD5 1 interaction is essential for DNA repair mechanisms and play significant role in tumor resistance to irradiation and chemotherapy treatments. Effective strategies to selectively interfere with BRCA2/RAD5 1 interaction in the context of treatment and chemoprevention of neoplastic diseases are described. (more...)

Human DNA polymerase e contains two apparent subunits of >200 and 55 kDa. Purified polymerase e was used to prepare mouse antiserum capable of recognizing different polymerase e subunits. A mouse monoclonal was identified that had specific recognition for the catalytic subunit of polymerase e (200 kDa). Reference: J Biol. Chem. (1995) v270, pp7799-7808. (more...)

Investigators at UCB have identified monoclonal cell lines, 2C3.11 and 5D5.8 that produce monoclonal IgG to the two subunits of human Ku autoantigen p70 and p85, repectively. Each antibody is able to recognize its particular antigen on denaturing immunoblots and each is able to immunoprecipitate the Ku autoantigen out of cell-free extracts. Ku autoantigen proteins are thought to be important in the process of repairing DNA strand breaks and recombining of chromosomes. (more...)

DNA at the end of chromosomes is maintained in a dynamic balance of loss and addition of simple sequence repeats (telomeres). Telomeric repeat addition is regulated and is catalyzed by the enzyme telomerase. Because normal somatic cells do not appear to highly express or require telomerase but cancer cells do highly express and require telomerase, molecules that can inhibit or effect telomerase activity are potential anti-cancer agents. The invention provides methods and compositions relating to a human telomerase and related nucleic acids, including four distinct human telomerase subunit proteins called p140, p105, p48, and p43 having human telomerase-specific activity. The proteins may be produced recombinantly from transformed host cells from the disclosed telomerase encoding nucleic acids or purified from human cells. Also included are human telomerase RNA components, as well as specific, functional derivatives thereof. The invention provides isolated telomerase hybridization probes and primers capable of specifically hybridizing with the disclosed telomerase gene, telomerase-specific binding agents such as specific antibodies, and methods of making and using the subject compositions in diagnosis, therapy and in the biopharmaceutical industry. (more...)

UC Berkeley scientists have identified several monoclonal antibodies (Mabs) that recognize several antigens in bubonic plague vaccine, including the Y. pestis Fraction I protein (F1) which is a universal indicator antigen in plague serology. These Mabs may be used to quantify antigens including F1 in commercially produced plague vaccine, for epitope mapping and determination of structure of these antigens as reagents for efficient immunoaffinity purification of the antigens, to screen recombinant hosts expressing cloned F1, and as reference reagents for analyzing the humoral immune response to plague infection and vaccination in animals and humans. (more...)

Regulated transcription requires specific interactions between activators bound to promoter sequences and other components of the RNA polymerase transcriptional machinery. These factors (TATA-binding protein Associated Factors [TAFs]) serve to mediate transcriptional activations UCB scientists have developed hybridomas producing monoclonal antibodies to: Drosophila TAFS 250, 150, 110, 80,60,40, 30alpha, 30beta and Human TAFS 250,130,100 References: The EMBO J (1993) v12 pp5303-9 Science (1994) v263 pp811-4 Genes & Dev. (1993) v7 pp2587-97 (more...)

Hybridoma cell lines that produce monoclonal antibodies with different specificities for the phenylurea herbicide diuron and its analogs. (more...)

Monoclonal antibodies that recognize phenanthrene, the chiral thiol and DNA adducts and metabolites of phenanthrene 9,10,-oxide, and structurally similar polynuclear aromatic K-region oxides (more...)

Specific polyclonal antibodies for assays of three thiocarbamate herbicides (molinate, thiobencarb, and EPTAM), and related compound (more...)

Prunus Necrotic Ringspot virus (PNRV) and its variants infect and have pathologic effects on a wide variety of stone fruit trees, apples, hops, and roses. This large economic impact necessitates a simple, sensitive, and highly reliable diagnostic test for these viruses in leaves, active and dormant buds, and other tissues. UC Berkeley researchers have developed 405 hybridoma lines specific for PNRV. To date, five of these have been investigated in depth. In initial tests by four laboratories, these antibodies reacted with a variety of PNRV isolates from California that commercially available antisera and monoclonal antibodies recognized more weakly or not at all. (more...)

This Invention employs monoclonal antibodies and direct gelling probes as molecular markers specific for gelling and nongelling parts of plant and bacterial carbohydrates. This invention describes a rapid assay which can detect these carbohydrates and analyze their composition in very small samples. The sample, such as ice cream or hamburger, is blotted onto a membrane, reacted with the molecular marker and then visualized with a marker detection system. Multiple samples can be analyzed rapidly and efficiently using this method. Gelling probes have been tested for several carbohydrates: alginate, pectate, carrageenan and agarose. These probes consist of short gelling subunits conjugated to markers. The direct gelling probes bind to the homologous carbohydrate in the sample on the blot only in the presence of the appropriate gelling ion or after a specific thermal change, depending on the specific carbohydrate system. The direct gelling probes can be identified on the filter paper similar to DNA probes. (more...)

Streptavidin-metallothionein chimeric proteins with biological recognition specificity in which the streptavidin moiety provides high affinity biotin binding and the metallothionein moiety provides a high affinity metal binding. The binding affinity of the streptavidin-metallothionein chimeric protein both for biotin and heavy metal ions allows specific incorporation into, conjugation with, or labelling of any biological material containing biotin with various heavy metal ions. (more...)

Investigators at University of California have developed a hybridoma cell line that produces monoclonal antibodies with specificity for the epsilon chain of the CD3 complex in the mouse, This antibody can be used to study the expression and function of CD3 on murine thymocytes. Reference: Allison et al. 1988. The T-Cell Receptor pp33-45. Havran et al. 1987. Nature 330:170-3 (more...)

Avermectins are a class of potent, broad-spectrum antibiotic and pesticidal substances that are used in a wide array of applications. Extraction and instrumental analysis methods for avermections are complex, lengthy and expensive, making immunoassay a desirable alternative. The University of California, Berkeley, at its Hybridoma Facility has developed monoclonal antibodies that have specificity to the following avermectins: Aver. B2A2 MAb that recognizes ivermectin, abamectin, but not avermectin A2 Aver. C4D6 MAb with preference for ivermectin Aver. B11C2 MAb that recognizes ivermectin, abamectin, and avermectin A2 Aver. C1A3 Similar to MAb B2A2 Aver. C5D6 Similar to MAb C4D6 Reference; Karu et al. Avermectins Detection with Monoclonal Antibodies 1990. ACS Symposium Series No. 442 Immunochemical Methods for Environmental Analysis. pp95-111. (more...)

This invention relates to the development of monoclonal antibodies that enable the detection of defects in neural development in Drosophila. The antibody shows strong specificity for axons with no staining of neuron cell bodies, the PNS, or other embryo tissues. Specificity is retained after glutaraldehyde treatment but is lost with periodate, indicating that the antibody likely recognizes a carbohydrate epitope. Published references; Seeger, M & et al. 1993. Mutations affecting growth cone guidence in Drosophila: genes necessary for guidance toward or away form the midline. Neuron. 10:409-26 Patel, N.H. 1994. Imaging neuronal subsets and other cell types in whole mount Drosphila embryos and larvae using antibody probes. In "Methods in Cell Biology, Vol 44. Drosophila melangaster: Practical Uses in Cell Biology", L.S.B. Goldstein and E. Fyrberg, eds. Academic Press, New York pp. 445-487. (more...)

In an effort to produce improved antibody reagents for non-muscle myosin II, investigators at University of California, Berkeley have developed mouse monoclonal antibodies against myosin II protein isolated form cytosol of rabbit gastric homogenates. Hybridoma clones were tested for the ability of their antibodies to recognize non-muscle myosin II from gastric fractions. Two clones were identified as producing good and specific antibodies against non-muscle myosin II protein. (more...)

Cell hybridomas were were generated from Listeria monocytogenes immunized CBA/J mice. Optimal immunity to the Gram-positive pathogen Listeria monocytogenes (LM) requires both CD8+ and CD4+ antigen-specific T cell responses. Understanding how CD4+ T cells function in an immune response to LM and how bacterial proteins are processed to peptide/MHC class II complexes in infected cells requires identification of these proteins. Using LacZ-inducible, LM-specific CD4+ T cells as probes, we identified two immunogenic LM proteins by a novel expression cloning strategy. The antigenic peptides contained within these proteins were defined by deletion analysis of the genes, and their antigenicity was confirmed with synthetic peptides. The nucleotide sequences of the genes showed that they encode previously unknown LM proteins that are homologous to surface proteins in other bacterial species. References: Sanderson et al. 1995. J. Exp. Med. 182:1751-7 Campell and Shastri 1998 J. Immunol 161:2339-47 (more...)

Minor histocompatibility (H) antigens elicit T cell responses and thereby cause chronic graft rejection and graft-vs.-host disease among MHC identical individuals. Although numerous independent H loci exist in mice of a given MHC haplotype, certain H antigens dominate the immune response and are thus of considerable conceptual and therapeutic importance. The H60 gene was isolated as a cDNA clone from the mouse strain BALB.B. This gene contains an antigenic peptide that elicits a strong cytotoxic T cell response when C57BL/6 mice are immunized with BALB.B spleen cells. References: Malarkannan et al. 1998. J Immunol. 161:3501-9. Karttunen et al. 1992. PNAS 89:6020-4. (more...)

Investigators at University of California have made a panel of hybridomas producing monoclonal antibodies to the ectodomain of murine ICOS from Syrian hamsters with Chinese hamster ovary cells (CHO) transfected with the murine ICOS gene. Two hybridoma lines were identified; 27A12 (Igm) and 151-9 (IgG). Ref: Dong, C, Juedes, AE, Temann, UA, Shresta, S, Allison, JP, Ruddle, NH, and Flavell, RA. 2001. ICOS co-stimulatory receptor is essential for T-cell activation and function. Nature. 409:97-101 (more...)

This invention concerns a hybridoma producing a rat (IgG2a, kappa) monoclonal antibody specific for the mouse NKG2 receptor subunits. The antibody reacts with three NKG2 subunits, NKG2A, NKG2C, and NKGE. These NKG2 subunits pair with the CD94 subunit, and are expressed on the surface of approximately 50% of NK cells and class I molecules. Ligand binding inhibits the NK cell when CD94 is paired with NKG2A, and probably activates the NK cell when CD94 is paired with NKG2C or E. Approximately half of NK cells do not express NKG2A, C, or E; these cells express CD94 on the surface in a form that does not bind ligand and is believed to be nonfunctional. Ref: Vance, RE, Jamieson, AM, and Raulet DH. 1999. recognition of the class Ib Molecule Qa-1b by Putative Activating Receptors CD94/NKG2C and CD94/NKG2E on Mouse Natural Killer Cells. J.Exp.Med 190:1801-12. (more...)

This invention concerns a hybridoma producing a mouse IgG1/kappa monoclonal antibody specific for the mouse Ly49F receptor. Mouse Ly49F is a homodimeric protein expressed on the surface of approximately 10% of NK cells, and a small fraction of T cells. Of T cells that express Ly49 receptors (approximately 15% of CD8 T cells), Ly49F is expressed weakly to the Dd classical class I molecule. The structure of the receptor suggests it may exert an inhibitory function. (more...)

This invention represents a hybridoma producing an hamster IgG monoclonal antibody specific for the mouse Ly49C, Ly49F, LY49H and Ly49I receptors. These receptors are homodimeric and are expressed by subsets f NK Cells and some T cells. This monoclonal reacts with all four receptors, resulting in reactivity with 90% of NK cells in some mouse strains. The Ly49 receptors bind to classical class I molecules. Ligand binding by Ly49C, F and I is expected to inhibit NK cell functions such as cytotoxicity. L:y49H, in contrast, is a stimulatory receptor on NK cells. The 14B11 antibody can stimulate killing by NK cells, probably because of its reactivity with Ly49H. Further detailed information was published; Corral, L. & et al. (1999) Hybridoma. 18:359-366 (more...)

This invention consists of a hybridoma cell line producing a rat (IgG2a, kappa) monoclonal antibody specific for the mouse CD94 receptor subunit. CD94 is expressed on the surface of all NK cells and some T cells. CD94 m when paired with an NKG2A, C or E subunit, binds the nonclassical Class I molecule Qa-1b associated with a peptide derived from classical class I molecules. Approximately half of NK cells do not express NKG2A, C, or E; these cells express CD94 on the surface in a form that does not bind Qa-1b and is believed to be nonfunctional. The utility of this monoclonal antibody was presented in the publication; Vance & et al. J. Exp Med. 1999 190:1801-12 (more...)