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Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: These are three ELISAs for detection of anti-HIV antibodies. These test were highly specific, sensitive and reproducible. They detect specific immunoglobulin Y against HIV peptides in the egg yolk of brown Leghorn layer hens. A modification of these methods were used to detect anti-HIV antibodies in Humans.
Proper citation: Angel Justiz-Vaillant 2020. ELISA for anti-HIV peptide antibodies in the egg yolks of brown Leghorn layer hens.. protocols.io dx.doi.org/10.17504/protocols.io.bjiqkkdw Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: This SpL ELISA can be used to detect specific antibodies in various animal species including human, mouse, rat, dog, rabbit, chicken, monkey, pig and hamster [1].1. De Chateau M, Nilson BH, Erntell M, Myhre E, Magnusson CG, Akerstrom B et al. On the interaction between protein L and immunoglobulins of various mammalian species. Scand J Immunol 1993; 37: 399−405
Proper citation: Angel Justiz-Vaillant 2020. Enzyme linked immunosorbent assay for investigating the binding of Protein-L to diverse immunoglobulins . protocols.io dx.doi.org/10.17504/protocols.io.bjpfkmjn Copy
Authors: Angel Justiz-Vaillant, Belkis Ferrer-Cosme
Group: University of the West Indies, [email protected]
Summary: MCP-1 is believed to play an important role in monocyte infiltration into tumor tissues. [1] MCP-1 plays an important role in the pathogenesis of atherosclerosis. There is considerable evidence that supports that monocytes containing MCPs and macrophages influence the growth of other cell types within the atherosclerotic lesion. [2]References1. Deshmane SL, Kremlev S, Amini S, Sawaya BE. Monocyte chemoattractant protein-1 (MCP-1): an overview. J Interferon Cytokine Res. 2009;29(6):313-326. doi:10.1089/jir.2008.00272. Lin J, Kakkar V, Lu X. Impact of MCP-1 in atherosclerosis. Curr Pharm Des. 2014;20(28):4580-4588. doi:10.2174/1381612820666140522115801
Proper citation: Angel Justiz-Vaillant, Belkis Ferrer-Cosme 2020. ELISA for measurement of monocyte chemoattractant protein-1 (MCP-1/CCL2) in human serum.. protocols.io dx.doi.org/10.17504/protocols.io.bj7fkrjn Copy
Authors: Angel Justiz-Vaillant, Norma McFarlane-Anderson
Group: University of the West Indies, [email protected]
Summary: This protocol was already used successfully to detect anti-HIV antibody in the serum of women with cervical dysplasia or cervical cancer in Jamaica, West Indies [1].Reference1. Justiz Vaillant A, Bazuaye PE, McFarlane-Anderson N, Smikle MF et al. Seroprevalence of Anti-HIV Antibodies in Women with Abnormal Pap Smears in Jamaica. British Journal of Medicine & Medical Research 2013, 3(4): 2197-2202.
Proper citation: Angel Justiz-Vaillant, Norma McFarlane-Anderson 2020. Indirect Enzyme Linked Immunosorbent Assay (ELISA) for Detection of Anti-HIV Antibodies in Human Serum. protocols.io dx.doi.org/10.17504/protocols.io.bkbmksk6 Copy
Authors: Angel Justiz-Vaillant, Norma McFarlane-Anderson
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant, Norma McFarlane-Anderson 2020. Universal sandwich enzyme linked immunosorbent assay for investigating protein-LG (SpLG) interactions with immunoglobulins using a SpA-HRP conjugate.. protocols.io dx.doi.org/10.17504/protocols.io.bjp4kmqw Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant 2020. Enzyme linked immunosorbent assay (ELISA) for determining the serum concentration of IL-33 in humans.. protocols.io dx.doi.org/10.17504/protocols.io.bjy9kpz6 Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells. They play essential roles in the activation and differentiation of immune cells, as well as proliferation, maturation, migration, and adhesion. They also have pro-inflammatory and anti-inflammatory properties. The primary function of interleukins is, therefore, to modulate growth, differentiation, and activation during inflammatory and immune responses. Interleukins consist of a large group of proteins that can elicit many reactions in cells and tissues by binding to high-affinity receptors in cell surfaces. CD4+T cells (Th2), NKT cells and mast cells synthesize IL-13. It acts on monocytes, fibroblasts, epithelial cells and B cells. The IL-13 significant effects are B-cell growth and differentiation, stimulates isotype switching to IgE. It causes increased mucus production by epithelial cells, increased collagen synthesis by fibroblasts and inhibits pro-inflammatory cytokine production. Also, IL-13 works together with IL-4 in producing biologic effects associated with allergic inflammation and in defense against parasites. [1]Reference1. Justiz Vaillant AA, Qurie A. Interleukin. In:StatPearls. Treasure Island (FL): StatPearls Publishing; June 12, 2019.
Proper citation: Angel Justiz-Vaillant 2020. ELISA for quantification of IL-13 in human serum.. protocols.io dx.doi.org/10.17504/protocols.io.bj3ckqiw Copy
Authors: Angel Justiz-Vaillant, Belkis Ferrer-Cosme
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant, Belkis Ferrer-Cosme 2020. ELISA for quantification of human C2 in serum or plasma.. protocols.io dx.doi.org/10.17504/protocols.io.bj73krqn Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells. They play essential roles in the activation and differentiation of immune cells, as well as proliferation, maturation, migration, and adhesion. They also have pro-inflammatory and anti-inflammatory properties. The primary function of interleukins is, therefore, to modulate growth, differentiation, and activation during inflammatory and immune responses. Interleukins consist of a large group of proteins that can elicit many reactions in cells and tissues by binding to high-affinity receptors in cell surfaces. Th9, Th2, Th17, mast cells, NKT cells, and regulatory T cells produce interleukin-9. It enhances T-cell survival, mast cell activation and synergy with erythropoietin.[1]Reference1. Justiz Vaillant AA, Qurie A. Interleukin. In:StatPearls. Treasure Island (FL): StatPearls Publishing; June 12, 2019.
Proper citation: Angel Justiz-Vaillant 2020. ELISA for quantification of IL-9 in human serum.. protocols.io dx.doi.org/10.17504/protocols.io.bj29kqh6 Copy
Authors: Norma McFarlane-Anderson
Group: University of the West Indies, [email protected]
Summary: This protocol was already used successfully to detect anti-HIV antibody in the serum of women with cervical dysplasia or cervical cancer in Jamaica, West Indies [1].Reference1. Justiz Vaillant A, Bazuaye PE, McFarlane-Anderson N, Smikle MF et al. Seroprevalence of Anti-HIV Antibodies in Women with Abnormal Pap Smears in Jamaica. British Journal of Medicine & Medical Research 2013, 3(4): 2197-2202.
Proper citation: Norma McFarlane-Anderson 2020. Indirect Enzyme Linked Immunosorbent Assay (ELISA) for Detection of Anti-HIV Antibodies in Human Serum. protocols.io dx.doi.org/10.17504/protocols.io.bjtkknkw Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells. They play essential roles in the activation and differentiation of immune cells, as well as proliferation, maturation, migration, and adhesion. They also have pro-inflammatory and anti-inflammatory properties. The primary function of interleukins is, therefore, to modulate growth, differentiation, and activation during inflammatory and immune responses. Interleukins consist of a large group of proteins that can elicit many reactions in cells and tissues by binding to high-affinity receptors in cell surfaces. They have both paracrine and autocrine function. Interleukins are also used in animal studies to investigate aspect related to clinical medicine.T cells and stem cells make IL-3. It functions as a multilineage colony-stimulating factor [1]CD4+T cells (Th2) synthesize IL-4, and it acts on both B and T cells. It is a B-cell growth factor and causes IgE and IgG1 isotype selection. It causes Th2 differentiation and proliferation, and it inhibits IFN gamma-mediated activation on macrophages. It promotes mast cell proliferation in vivo. [1]Reference1. Justiz Vaillant AA, Qurie A. Interleukin. In:StatPearls. Treasure Island (FL): StatPearls Publishing; June 12, 2019.
Proper citation: Angel Justiz-Vaillant 2020. ELISA for quantification of IL-4 in human serum.. protocols.io dx.doi.org/10.17504/protocols.io.bj24kqgw Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells. They play essential roles in the activation and differentiation of immune cells, as well as proliferation, maturation, migration, and adhesion. They also have pro-inflammatory and anti-inflammatory properties. The primary function of interleukins is, therefore, to modulate growth, differentiation, and activation during inflammatory and immune responses. Interleukins consist of a large group of proteins that can elicit many reactions in cells and tissues by binding to high-affinity receptors in cell surfaces. Various phagocytes and epithelial cells synthesize Interleukin-34 (IL-34). It enhances IL-6 production and participates in the differentiation and development of antigen-presenting cells including microglia. [1]Reference1. Justiz Vaillant AA, Qurie A. Interleukin. In:StatPearls. Treasure Island (FL): StatPearls Publishing; June 12, 2019.
Proper citation: Angel Justiz-Vaillant 2020. ELISA for quantification of IL-34 in human serum.. protocols.io dx.doi.org/10.17504/protocols.io.bj36kqre Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: 1. IBP-HRP conjugates can be prepared in the laboratory by the periodate method.2. The Human IgG used can be commercially available of prepared in the laboratory from human sera by affinity chromatography (using an SpG-agarose column).3. SpG-agarose column are widely commercially prepared.4. PURE-1A antibody purification kit will be discontinued. However there are others product for this purpose as Montage Antibody Purification Kit with PROSEP-G media.
Proper citation: Angel Justiz-Vaillant 2020. Binding properties of Human IgG to immunoglobulin-binding proteins tested by double immunodiffusion (Ouchterlony) technique.. protocols.io dx.doi.org/10.17504/protocols.io.bjskkncw Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Avian IgY is structurally and genetically different from its mammalian counterpart IgG. This reagent can be used in ELISA, Western blotting and Dot blot to detect specific chicken IgY to infectious agents or immune system proteins [1]. IgY can be isolated by the Polson Method [2] or the water dilution method [3]. Chicken IgY can crossed-react with IgY from diverse bird species [4]. Similarly avian IgY can be isolated from their egg yolk using the same methodology with modifications that was developed for the chicken IgY isolation [5].Reference1. Larsson A, Bålöw RM, Lindahl TL, Forsberg PO. Chicken antibodies: taking advantage of evolution--a review. Poult Sci. 1993;72(10):1807-1812. doi:10.3382/ps.07218072.Polson A (1990) Isolation of IgY from the yolks of eggs by a chloroform polyethylene glycol procedure. Immunol Invest 19: 253-258. 3. Nilsson E, Hanrieder J, Bergquist J, Larsson A. Proteomic characterization of IgY preparations purified with a water dilution method.J Agric Food Chem. 2008;56(24):11638-11642. doi:10.1021/jf802626t4. Vaillant AJ, McFarlane-Andersonv N, Wisdom B, Mohammed W, Vuma S, et al. (2013) Immunoglobulin-binding Bacterial Proteins (IBP) Conjugates and their Reactivity with Immunoglobulin in Enzyme-Linked Immunosorbent Assays (ELISA). J Anal Bioanal Tech 4: 175. doi:10.4172/2155-9872.1000175 5. : Justiz Vaillant AA, Ramirez N, Cadiz A, Ferrer B, Akpaka P, et al. (2013) Separation and Reactivity of Avian Immunoglobulin Y. J Chromat Separation Techniq 4: 173. doi:10.4172/2157-7064.1000173
Proper citation: Angel Justiz-Vaillant 2020. Preparation of horseradish peroxidase (HRP) conjugated to chicken anti-IgY . protocols.io dx.doi.org/10.17504/protocols.io.bjmvkk66 Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant 2020. Binding properties of immunoglobulin-binding protein by double immunodiffusion (Ouchterlony) technique. . protocols.io dx.doi.org/10.17504/protocols.io.bjqdkms6 Copy
Authors: Angel Justiz-Vaillant, Belkis Ferrer-Cosme
Group: University of the West Indies, [email protected]
Summary: IFN-γ is a key player in driving cellular immunity. It is capable of orchestrating numerous and vital protective functions to enhance immune responses in cancers and infectious diseases. It exhibits its immunomodulatory effects by enhancing antigen processing and presentation,, inducing an anti-viral state, increasing leukocyte trafficking, boosting the anti-microbial properties of immune cells and by affecting cellular proliferation and apoptosis. [1]ReferenceKak G, Raza M, Tiwari BK. Interferon-gamma (IFN-γ): Exploring its implications in infectious diseases.Biomol Concepts. 2018;9(1):64-79. Published 2018 May 30. doi:10.1515/bmc-2018-0007
Proper citation: Angel Justiz-Vaillant, Belkis Ferrer-Cosme 2020. ELISA for quantification of interferon gamma (IFN-γ) in human serum or plasma.. protocols.io dx.doi.org/10.17504/protocols.io.bj68krhw Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: SpLG comprises of 4 Ig-binding domains of SpL and 2 IgG Fc-binding SpG domains [1]. This hybrid molecule was found to bind many intact human Ig molecules and Ig fragments [1]. It proved a powerful tool for the binding, detection and purification of antibodies [2].It was reported that chimeric SpLG was a potent mitogen for mouse splenic B cells, and induced cell differentiation and the production of immunoglobulins. Inhibition experiments demonstrated that the Ig-binding capacity of both SpG and SpL in the chimeric molecule are independent of each other. It was also shown that SpLG selectively absorbed Igs present in the sera of humans, rabbits, mice and rats [1-3]. The preparation of SpLG-HRP by the periodate method is a novel application [4].References1. Justiz-Vaillant AA, Akpaka PE, McFarlane-Anderson N, Smikle MF. Comparison of techniques of detecting immunoglobulin-binding protein reactivity to immunoglobulin produced by different avian and mammalian species.West Indian Med J. 2013;62(1):12-20.2. Kihlberg BM, Sjöbring U, Kastern W, Björck L. Protein LG: a hybrid molecule with unique immunoglobulin binding properties. J Biol Chem 1992, 267(35): 25583-8.3. Justiz-Vaillant AA, McFarlane-Anderson N, and Smikle M. “Bacterial Immunoglobulin (Ig)-Receptors: Past and Present Perspectives.” American Journal of Microbiological Research, vol. 5, no. 2 (2017): 44-50. doi: 10.12691/ajmr-5-2-4.4. Vaillant AJ, McFarlane-Andersonv N, Wisdom B, Mohammed W, Vuma S, et al. (2013) Immunoglobulin-binding Bacterial Proteins (IBP) Conjugates and their Reactivity with Immunoglobulin in Enzyme-Linked Immunosorbent Assays (ELISA). J Anal Bioanal Tech 4: 175. doi:10.4172/2155-9872.1000175.
Proper citation: Angel Justiz-Vaillant 2020. Preparation of a protein-LG conjugated to horseradish peroxidase by the periodate method.. protocols.io dx.doi.org/10.17504/protocols.io.bjk3kkyn Copy
Authors: Angel Justiz-Vaillant, Suzette E. Curtello
Group: University of the West Indies, [email protected]
Summary: Salmonella are motile, flagellated rod-shaped Zoonotic pathogens which may survive with or without oxygen and do not absorb crystal violet stain.They are decolourised by alcohol due to their outer lipopolysaccharide membrane and thin peptidoglycan layer.They belong to the family Enterobacteriaceae and is implicated with typhoid fever and food-borne illnesses.This pathogen is associated with intestinal disease which may become fatal and has negatively impact the health of individuals and various economies globally.The poultry industry is most impacted and vulnerable to the onslaught of this pernicious microbe.The Lipopolysaccharide Somatic O antigen, flagellar H and virulent V antigenic structure determines the serotype designate of Salmonella species [1]. A wide range of antisera is used in the location of the O antigen associated with the carbohydrate constituent of Lipopolysaccharide and H protein antigen linked with the flagella of the bacteria.Salmonellosis is an inflammation of the intestinal mucosal lining by Salmonella bacteria infiltration resulting in painful symptoms.Both man and animal are vulnerable.Salmonellosis is caused by ingestion of Salmonella with food (Wall et al., 1994).The main route of transmission of Salmonella microbe is fecal-oral. Ingestion of small concentration of Salmonella such as 6 cells with contaminated food or water will lead to salmonellosis infection [2].References1. Varma JK, Marcus R, Stenzel SA, et al. Highly resistant Salmonella Newport-MDRAmpC transmitted through the domestic US food supply: a FoodNet case-control study of sporadic Salmonella Newport infections, 2002-2003.J Infect Dis. 2006;194(2):222-230. doi:10.1086/505084.2. Chen W, Martinez G, Mulchandani A. Molecular beacons: a real-time polymerase chain reaction assay for detecting Salmonella.Anal Biochem. 2000;280(1):166-172. doi:10.1006/abio.2000.4518
Proper citation: Angel Justiz-Vaillant, Suzette E. Curtello 2020. Salmonella Typhimurium isolation from specimens. protocols.io dx.doi.org/10.17504/protocols.io.bjjdkki6 Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: The peroxidase-labeled anti-chicken IgY conjugate cross-reacts with many IgY present in the egg of many and diverse avian species. References1. Justiz Vaillant AA, Ramirez N, Cadiz A, Ferrer B, Akpaka P, et al. (2013) Separation and Reactivity of Avian Immunoglobulin Y. J Chromat Separation Techniq 4: 173. doi:10.4172/2157-7064.1000173
Proper citation: Angel Justiz-Vaillant 2020. Direct ELISA for investigating the binding of peroxidase-labeled anti-chicken IgY conjugate with avian immunoglobulins. protocols.io dx.doi.org/10.17504/protocols.io.bjxykppw Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Chicken immunization with peptides is inefective if only just the peptides are being inoculated. However, to make the immune response effective the fragment 254-274 of HIV-1 was conjugated with a carrier protein (KLH) that produced a critical immune response, assessed by ELISA, Immunoblot analysis and dot blot [1-4]. The Polson method (1990) can be used effectively to separate the IgY antibody from the egg yolk of immunized chickens [5].References1. Schade R, Pfister C, Italatsch R, Henklien P (1991) Polyclonal antibodies from chicken egg yolk-an alternative to the production of mammalian IgG type antibodies in rabbits. ATLA 19: 403-419. 2. Justiz Vaillant AA, Akpaka PE, Smikle M, McFarlane-Anderson N (2012) In vitro Inhibition of Staphylococcus aureus Isolates by Anti-Anti-Idiotypic Antibodies to Staphylococcal Protein (SpA). J Vaccines Vaccin 3: 127-131.3. Collins AM, Roberton DM, Hosking CS, Flannery GR (1991) Oral immunization with xenogeneic antibodies stimulates the production of systemic and mucosal anti-idiotypic antibodies. Immunology 73: 388-393. 4. Ho DD, Kaplan JC, Rackauskas IE, Gurney ME (1988) Second conserved domain of gp120 is important for HIV infectivity and antibody neutralization. Science 239: 1021-1023. 5. Polson A (1990) Isolation of IgY from the yolks of eggs by a chloroform polyethylene glycol procedure. Immunol Invest 19: 253-258.
Proper citation: Angel Justiz-Vaillant 2020. Chicken immunization with Keyhole limpet hemocynin (KLH)-gp120 fragment (254-274) conjugate raises anti-KLH antibodies in egg yolks.. protocols.io dx.doi.org/10.17504/protocols.io.bjvtkn6n Copy
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