Creative Diagnostics
 
 
Nanobodies are antibody-derived therapeutic proteins that contain the unique structural and functional properties of naturally-occurring heavy-chain antibodies. The Nanobody technology was originally developed following the discovery that camelidae (camels and llamas) possess fully functional antibodies that lack light chains. These heavy-chain antibodies contain a single variable domain (VHH) and two constant domains (CH2 and CH3). Importantly, the cloned and isolated VHH domain is a perfectly stable polypeptide harbouring the full antigen-binding capacity of the original heavy-chain antibody.
 
Creative Diagnostics is focused on the discovery and development of Nanobodies, for a range of serious human diseases including inflammation, haematology, oncology and pulmonary disease.
 
ALB
 
Full Name
Acidovorax citrulli
 
Background
Acidovorax citrulli (formerly A. avenae subsp. citrulli) is a Gram-negative, biotrophic bacterium causes seedling blight and bacterial fruit blotch (BFB) of cucurbits. On the basis of carbon source utilization, DNA-fingerprinting profiles, whole-cell fatty-acid composition utilization and pathogenicity assays, A. citrulli is divided in to two distinct groups. The group I strains are mainly associated with non-watermelon plants (mainly melon), while group II includes strains that were mainly isolated from watermelon.
 
CCL11
 
Full Name
chemokine (C-C motif) ligand 11
 
Background
Chemokine (C-C motif) ligand 11 (CCL11) is a small cytokine belonging to the CC chemokine family that is also known as eotaxin-1. CCL11 selectively recruits eosinophils by inducing their chemotaxis, and therefore, is implicated in allergic responses. The effects of CCL11 are mediated by its binding to a G-protein-linked receptor known as a chemokine receptor. Chemokine receptors for which CCL11 is a ligand include CCR2, CCR3 and CCR5. However, it has been found that eotaxin-1 (CCL11) has high degree selectivity for its receptor, such that they are inactive on neutrophils and monocytes, which do not express CCR3.[5] The gene for human CCL11 (scya11) is encoded on three exons and is located on chromosome 17.
 
CCL3
 
Full Name
chemokine (C-C motif) ligand 3
 
Background
CCL3, also known as Macrophage inflammatory protein-1α (MIP-1α), is a cytokine belonging to the CC chemokine family that is involved in the acute inflammatory state in the recruitment and activation of polymorphonuclear leukocytes (Wolpe et al., 1988). Sherry et al. (1988) demonstrated 2 protein components of MIP1, called by them alpha and beta.
 
CCL5
 
Full Name
chemokine (C-C motif) ligand 5
 
Background
It is also known as RANTES (Regulated upon Activation, Normal T-cell Expressed, and Secreted). CCL5 is an 8kDa protein classified as a chemotactic cytokine or chemokine. CCL5 is chemotactic for T cells, eosinophils, and basophils, and plays an active role in recruiting leukocytes into inflammatory sites. With the help of particular cytokines (i.e., IL-2 and IFN-γ) that are released by T cells, CCL5 also induces the proliferation and activation of certain natural-killer (NK) cells to form CHAK (CC-Chemokine-activated killer) cells.[2] It is also an HIV-suppressive factor released from CD8+ T cells. This chemokine has been localized to chromosome 17 in humans. RANTES was first identified in a search for genes expressed late (3–5 days) after T cell activation. It was subsequently determined to be a CC chemokine and expressed in more than 100 human diseases. RANTES expression is regulated in T lymphocytes by Kruppel like factor 13 (KLF13). RANTES, along with the related chemokines MIP-1alpha and MIP-1beta, has been identified as a natural HIV-suppressive factor secreted by activated CD8+ T cells and other immune cells.
 
CD28
 
Full Name
CD28 CD28 molecule
 
CEACAM1
 
Full Name
CEACAM1 carcinoembryonic antigen-related cell adhesion molecule 1 (biliary glycoprotein)
 
CTLA4
 
Full Name
Ctla4 cytotoxic T-lymphocyte-associated protein 4
 
Background
CD152 is a 33 kD member of the immunoglobulin superfamily also known as CTLA-4 or Ly-56. It is expressed on activated T and B lymphocytes. CD152 is similar to CD28 in amino acid sequence, structure, and genomic organization and these two receptors share common B7 family counter-receptors (B7-1, B7-2). Whereas CD28 delivers a costimulatory signal in T cell activation, CTLA-4 negatively regulates cellmediated immune responses. CD152 is thought to play a role in the induction and maintenance of immunological tolerance, as well as the development of protective immunity and thymocyte regulation.
 
CXCR4
 
Full Name
chemokine (C-X-C motif) receptor 4
 
Background
This gene encodes a CXC chemokine receptor specific for stromal cell-derived factor-1. The protein has 7 transmembrane regions and is located on the cell surface. It acts with the CD4 protein to support HIV entry into cells and is also highly expressed in breast cancer cells. Mutations in this gene have been associated with WHIM (warts, hypogammaglobulinemia, infections, and myelokathexis) syndrome. Alternate transcriptional splice variants, encoding different isoforms, have been characterized.
 
EGFR
 
Full Name
epidermal growth factor receptor
 
Background
The epidermal growth factor receptor (EGFR; ErbB-1; HER1 in humans) is the cell-surface receptor for members of the epidermal growth factor family (EGF-family) of extracellular protein ligands. The epidermal growth factor receptor is a member of the ErbB family of receptors, a subfamily of four closely related receptor tyrosine kinases: EGFR (ErbB-1), HER2/c-neu (ErbB-2), Her 3 (ErbB-3) and Her 4 (ErbB-4). Mutations affecting EGFR expression or activity could result in cancer. Epidermal Growth Factor was discovered by Stanley Cohen of Vanderbilt University along with Rita Levi-Montalcini for which both received the Nobel prize in Physiology or Medicine in 1986.
 
ERBB2
 
Full Name
v-erb-b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog (avian)
 
Background
HER2/neu (also known as ErbB-2) stands for Human Epidermal growth factor Receptor 2 and is a protein giving higher aggressiveness in breast cancers. It is a member of the ErbB protein family, more commonly known as the epidermal growth factor receptor family. HER2/neu has also been designated as CD340 (cluster of differentiation 340) and p185. It is encoded by the ERBB2 gene. Defects in ERBB2 are associated with gastric cancer [MIM:137215]; also known as hereditary familial diffuse gastric cancer (HDGC). Defects in ERBB2 are involved in the development of glioma (GLM) [MIM:137800]. Gliomas are central nervous system neoplasms derived from glial cells and comprise astrocytomas, glioblastoma multiforme, oligodendrogliomas, and ependymomas.
 
FGFR1
 
Full Name
fibroblast growth factor receptor 1
 
Background
Fibroblast growth factor receptor 1 (FGFR1), also known as basic fibroblast growth factor receptor 1, fms-related tyrosine kinase-2 / Pfeiffer syndrome, and CD331, is a receptor tyrosine kinase whose ligands are specific members of the fibroblast growth factor family. FGFR1 has been shown to be associated with Pfeiffer syndrome.
 
GFP
 
Full Name
Green Fluorescent Protein
 
Background
Green fluorescence protein (GFP) is a 27 kDa protein derived from the jellyfish Aequorea victoria, which emits green light (emission peak at a wavelenth of 509 nm) when excited by blue light (excitation peak at a wavelenth of 395 nm). GFP has become an invaluable tool in cell biology research, since its intrinsic fluorescence can be visualized in living cells. GFP fluorescence is stable under fixation conditions and suitable for a variety of applications. GFP has been widely used as a reporter for gene expression, enabling researchers to visualize and localize GFP-tagged proteins within living cells without the need for chemical staining. Other applications of GFP include assessment of protein protein interactions through the yeast two hybrid system and measurement of distance between proteins through fluorescence energy transfer (FRET) protocols. GFP technnology has considerably contributed to a greater understanding of cellular physiology. YFP differs from GFP due to a mutation at T203Y; antibodies raised against full-length GFP should also detect YFP and other variants.
 
GST
 
Full Name
Glutathione-S-Transferase
 
Background
Enzymes of the glutathione S-transferase (GST) family are composed of many cytosolic, mitochondrial, and microsomal (now designated as MAPEG) proteins. GSTs are present in eukaryotes and in prokaryotes, where they catalyze a variety of reactions and accept endogenous and xenobiotic substrates.
 
HIF1A
 
Full Name
hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor)
 
Background
Hypoxia, a condition of low tissue O2 concentration, plays an important role in normal physiological processes and tumor formation. Under hypoxic conditions mammalian cells up regulate the expression of hypoxic genes, including induction of angiogenesis and a switch to anaerobic metabolism, in order to survive. HIF-1 (Hypoxia Inducible Factor-1) is one of the key regulators of the transcriptional response to oxygen deprivation (1). HIF-1 is composed of two subunits, HIF-1alphaand HIF-1beta also known as aryl hydrocarbon receptor nuclear translocator (ARNT)) that are members of the basic helix-loop-helix (bHLH) Per-Arnt-Sim (PAS) (bHLH-PAS) family of transcription factors. HIF-1 is essential for angiogenesis, embryonic development, and is associated with tumor progression, erythropoiesis, vascular development/remodeling, vasodilation, and glucose/energy metabolism. The over expression of HIF-1alphahas been demonstrated in many common human cancers including prostate and breast, in which HIF-1alpha levels are associated with increase vascularitry and tumor progression. Besides physiological hypoxia, genetic abnormalities frequently detected in human cancers, such as loss of function mutations (Von Hippel-Lindau, p53, and PTEN), are associated with induction of HIF1 activity and expression of HIF-1-inducible genes (1).
 
IGHE
 
Full Name
Immunoglobulin heavy constant epsilon
 
IL12B
 
Full Name
Il12b interleukin 12b
 
Background
IL12 is secreted by peripheral lymphocytes after induction. It is produced mainly by B-cells and to a lesser extent by T-cells. The most powerful inducers of IL12 are bacteria, bacterial products, and parasites. IL12 is produced after stimulation with Phorbol esters or Calcium ionophore by human B-lymphoblastoid cells. The IL12 receptor,CD212, appears to be a single protein of approximately 110 kDa. Up to 1000-9000 high affinity IL12 receptors/cell are expressed on peripheral blood mononuclear cells activated by various mitogens for T-cells or by IL2. IL12 receptors are present on activated T-cells expressing CD4 and CD8 and on activated CD56(+) natural killer cells. Resting peripheral blood mononuclear cells, tonsillar B-cells, or tonsillar B-cells activated by anti-IgM/Dx, anti-IgM/Dx + IL2, or SAC + IL2 do not express the receptor. Human IL12 is not active in murine lymphocytes. Hybrid heterodimers consisting of murine p35 and human p40 subunits retain bioactivity on murine cells; however, the combination of human p35 and murine p40 is completely inactive on murine cells. Murine IL12 is active on both murine and human lymphocytes.
 
IL13
 
Full Name
interleukin 13
 
Background
This gene encodes an immunoregulatory cytokine produced primarily by activated Th2 cells. This cytokine is involved in several stages of B-cell maturation and differentiation. It up-regulates CD23 and MHC class II expression, and promotes IgE isotype switching of B cells. This cytokine down-regulates macrophage activity, thereby inhibits the production of pro-inflammatory cytokines and chemokines. This cytokine is found to be critical to the pathogenesis of allergen-induced asthma but operates through mechanisms independent of IgE and eosinophils. This gene, IL3, IL5, IL4, and CSF2 form a cytokine gene cluster on chromosome 5q, with this gene particularly close to IL4.
 
IL23A
 
Full Name
interleukin 23, alpha subunit p19
 
Background
Interleukin-23 (IL-23) is a heterodimeric cytokine consisting of two subunits, one called p40, which is shared with another cytokine, IL-12, and another called p19 (the IL-23 alpha subunit). IL-23 is an important part of the inflammatory response against infection. It promotes upregulation of the matrix metalloprotease MMP9, increases angiogenesis and reduces CD8+ T-cell infiltration. Recently, IL-23 has been implicated in the development of cancerous tumors. In conjunction with IL-6 and TGF-β1, IL-23 stimulates naive CD4+ T cells to differentiate into a novel subset of cells called Th17 cells, which are distinct from the classical Th1 and Th2 cells. Th17 cells produce IL-17, a proinflammatory cytokine that enhances T cell priming and stimulates the production of proinflammatory molecules such as IL-1, IL-6, TNF-alpha, NOS-2, and chemokines resulting in inflammation. Knockout mice deficient in either p40 or p19, or in either subunit of the IL-23 receptor (IL-23R and IL12R-β1) develop less severe symptoms of multiple sclerosis and inflammatory bowel disease highlighting the importance of IL-23 in the inflammatory pathway.
 
IL6
 
Full Name
Interleukin 6
 
Background
This gene encodes a cytokine that functions in inflammation and the maturation of B cells. In addition, the encoded protein has been shown to be an endogenous pyrogen capable of inducing fever in people with autoimmune diseases or infections. The protein is primarily produced at sites of acute and chronic inflammation, where it is secreted into the serum and induces a transcriptional inflammatory response through interleukin 6 receptor, alpha. The functioning of this gene is implicated in a wide variety of inflammation-associated disease states, including suspectibility to diabetes mellitus and systemic juvenile rheumatoid arthritis.
 
IL6R
 
Full Name
IL6R interleukin 6 receptor
 
Background
The IL-6 receptor is a complex of 2 molecules, CD126 and gp130 (CD130).20 The alpha-chain CD126 is a glycoprotein that contains the ligand-binding site.The soluble form of CD126 that is generated by both normal and myeloma cells23 can bind IL-6 and mediate signal transduction through membrane-bound CD130.
 
KLK3
 
Full Name
kallikrein-related peptidase 3
 
Background
Prostate-specific antigen (PSA) is a protein produced by the cells of the prostate gland.PSA is present in small quantities in the serum of men with healthy prostates, but is often elevated in the presence of prostate cancer and in other prostate disorders. A blood test to measure PSA is considered the most effective test currently available for the early detection of prostate cancer, but this effectiveness has also been questioned, since it has a positive predictive value of only about 35%.
 
MMP12
 
Full Name
matrix metallopeptidase 12 (macrophage elastase)
 
Background
Human macrophage elastase (MMP-12) is a member of the family of matrix metalloproteinases (MMPs) that plays, like other members of the family, an important role in inflammatory processes contributing to tissue remodeling and destruction. In particular, a prominent role of MMP-12 in the destruction of elastin in the lung alveolar wall and the pathogenesis of emphysema has been suggested (1). MMP-12 expressed mainly in alveolar macrophages has been identified in the mouse lung as the main destructive agent associated with cigarette smoking, which gives rise to emphysema, both directly via elastin degradation and indirectly by disturbing the proteinase/antiproteinase balance via inactivation of the alpha1-proteinase inhibitor (alpha1-PI), the antagonist of the leukocyte elastase (2). Because elastin represents a critical component of aortic wall structure and a matrix substrate for metalloelastases, MMP-12 may have a direct and singular role in the pathogenesis of aortic aneurysms (3).
 
PDCD1
 
Full Name
Pdcd1 programmed cell death 1
 
Background
CD279 is a 50-55 kD immunoglobulin superfamily member, also known as programmed death-1 (PD-1). PD-1 is expressed on a subset of CD4-CD8-thymocytes, and on activated T and B cells. PD-1 is thought to be involved in lymphocyte clonal selection and peripheral tolerance. The PD-1 ligands, PD-L1 (also known as B7-H1) and PD-L2 (B7-DC), are members of the B7 immunoglobulin superfamily.
 
TNFSF11
 
Full Name
tumor necrosis factor (ligand) superfamily, member 11
 
Background
This gene encodes a member of the tumor necrosis factor (TNF) cytokine family which is a ligand for osteoprotegerin and functions as a key factor for osteoclast differentiation and activation. This protein was shown to be a dentritic cell survival factor and is involved in the regulation of T cell-dependent immune response. T cell activation was reported to induce expression of this gene and lead to an increase of osteoclastogenesis and bone loss. This protein was shown to activate antiapoptotic kinase AKT/PKB through a signaling complex involving SRC kinase and tumor necrosis factor receptor-associated factor (TRAF) 6, which indicated this protein may have a role in the regulation of cell apoptosis. Targeted disruption of the related gene in mice led to severe osteopetrosis and a lack of osteoclasts. The deficient mice exhibited defects in early differentiation of T and B lymphocytes, and failed to form lobulo-alveolar mammary structures during pregnancy. Two alternatively spliced transcript variants have been found.
 
VWF
 
Full Name
von Willebrand factor
 
Background
The glycoprotein encoded by this gene functions as both an antihemophilic factor carrier and a platelet-vessel wall mediator in the blood coagulation system. It is crucial to the hemostasis process. Mutations in this gene or deficiencies in this protein result in von Willebrand's disease. An unprocessed pseudogene has been found on chromosome 22.
 
 

 

Nanobodies

Nanobodies are recombinant, single-domain, variable fragments of camelid heavy chain-only antibodies (~95 kDa), which are able to bind selectively to a specific antigen. Typically, nanobodies are the variable domain alone of heavy chain antibodies (i.e. VHH) with approximate molecular weight of 12-15 kDa and are regarded as the smallest naturally derived antigen-binding fragment.



 

 

Advantages of Nanobodies:

1. Nanobody is easier to be operated.

2. Higher hydrophilicity contributes to higher solubility of nanobodies.

3. Small molecular weight, which is only 15 kD, and therefore has good tissue penetration.

4. High homology with human and a small molecular weight lead to the weak immunogenicity of the human body.

 

 

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