Immunoglobulin G-Encapsulated Gold Nanoclusters as Fluorescent Tags for Dot-Blot Immunoassays

Klebsiella pneumoniae is the second most common cause of community infection and nosocomial infections due to Gram-negative bacteria. These bacteria are able to induce the onset of immune response, especially humoral immune response.Humoral immunity acts through the activation of B cells that produce antibodies. Antibodies, especially IgG, will cause encapsulated bacteria such as Klebsiella pneumoniae to be better phagocytosed. The purpose of this study was to determine the IgG response to hemagglutinin protein pili K. pneumoniae 12.8 kDa. The method used in this research is Immunoblotting method with western blot and dot blot. The primary antibodies used for the western blot and dot blot tests were isolated from BALB / C mice serum induced with the subcutaneous pili K. pneumoniae 12.8 kDa protein. To get the standard in assessing the results of dot blot were used Corel Photo-paint X6. The semi-quantitative result of dot blot was obtained with the strongest reaction of the antibody dilution at 1/100 while the antigen dilution titer at 1 / 10.000. Results from western blotting showed a positive reaction of the pili protein subunit with a molecular weight of 128.1 kDa, 114.4 kDa, 64.9 kDa, 31.1 kDa, 27.7 kDa, 24.8 kDa, 20.9 kDa, 12.8 kDa, and 10 kDa. The conclusion of this study is the immunization of hemagglutinin pili K. pneumoniae 12.8 kDa subcutaneously capable of inducing the formationof Immunoglobulin G in BALB / C mice.

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Identification of novel pig and human immunoglobulin G-binding proteins and characterization of the binding regions of enolase from Streptococcus suis serotype 2

10.21203/rs.3.rs-23090/v2 ◽

2020 ◽

Author(s):

Quan Li ◽

Yang Fu ◽

Genglin Guo ◽

Zhuohao Wang ◽

Wei Zhang

Keyword(s):

Western Blot ◽

Immunoglobulin G ◽

Binding Proteins ◽

Streptococcus Suis ◽

Factor H ◽

Surface Proteins ◽

Host Immune System ◽

Dot Blot ◽

Binding Regions ◽

Far Western Blot

Abstract Streptococcus suis, a major emerging pathogen in swine and humans, expresses immunoglobulin G (IgG)-binding proteins (IBPs), which contribute to the ability of organism to evasion of host immune system. The objective of this study was to identify novel pig IgG (pIgG) and human IgG (hIgG)-binding proteins and characterize the binding regions of enolase from Streptococcus suis serotype 2 (S. suis 2). Here, four pIgG-binding proteins (pIBPs) and five hIgG-binding proteins (hIBPs) were identified from S. suis 2 surface proteins by 2D-Far-western blot assays. All the newly captured proteins were expressed and further confirmed their binding activity to pIgG or hIgG by Far-western blot and dot blot. In addition to previously identified factor H, fibronectin, collagen, fibrinogen, plasminogen and laminin, we also found that both pIgG and hIgG can specifically interact with enolase. Binding assays indicated that interactions of S. suis 2 enolase with pIgG and hIgG is primarily mediated by the enolase C-terminal portion (Enolase-C, a.a. 142-432). We found that hIgG exhibited stronger binding ability to Enolase-C than pIgG. Further analysis of the C-terminal regions of enolase (Enolase-C1 and Enolase-C2) suggested that the C-terminus possessed two different binding domains with distinct host IgG proteins. Strikingly, we confirmed that pIgG interacted with the Enolase-C1 (a.a. 142-271) and hIgG interacted with the Enolase-C2 (a.a. 271-432). These observations of enolase provide interesting insights in the pathogenesis of S. suis infection.

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Hypogalactosylation of immunoglobulin G (IgG) of gingival fluid and saliva at the patient with periodontal disease

Stomatoloski glasnik Srbije ◽

10.2298/sgs0601007s ◽

2006 ◽

Vol 53 (1) ◽

pp. 7-16

Author(s):

Gordana Stefanovic ◽

Dragana Ciric ◽

Vesna Ilic ◽

Gavrilo Brajovic ◽

Sonja Petrovic ◽

...

Keyword(s):

Periodontal Disease ◽

Immunoglobulin G ◽

Clinical Picture ◽

Igg Subclasses ◽

Dot Blot ◽

Periodontal Status ◽

Gingival Fluid ◽

Control Samples

Changed glycosylation of immunoglobulin G (IgG), above all, the expression of thermal galactose, influence to numerous functions of those immunoglobulin and correlate with the inflammatory level in a number of diseases. Aim: This work analyses the distribution of IgG subclasses and the content of thermal galactose in them, in saliva and gingival fluid of the patients with periodontal disease and different gum inflammatory level. Materials and methods: It was used saliva and gingival fluid of 30 adults with clinical picture of periodontal disease and 20 persons with healthy periodontium. The qualification of IgG was done by ?dot-blot? procedure and the, and thermal galactose was determined by lectin immunoblot procedure. Results: The division of IgG subclasses in both fluids was different in the patients with periodontal disease and in control samples. In saliva and gingival fluid of the diseased quantitatively dominated IgG2 subclasses, independently from periodontal status. In IgG of both fluids, thermal galactose was exprimated at the healthy periodontium persons (control) and with the person with initial periodontal disease, while at the person with increased periodontal disease the expression of this saccharide wasn?t registered in neither of fluids. Conclusion: The results showed that there is a shift towards hypogalactosyled IgG glikoforms during the process of gum inflammation at the periodontal disease patients. .

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Identification of novel pig and human immunoglobulin G-binding proteins and characterization of the binding regions of enolase from Streptococcus suis serotype 2

10.21203/rs.3.rs-23090/v1 ◽

2020 ◽

Author(s):

Quan Li ◽

Yang Fu ◽

Genglin Guo ◽

Zhuohao Wang ◽

Wei Zhang

Keyword(s):

Western Blot ◽

Immunoglobulin G ◽

Binding Proteins ◽

Streptococcus Suis ◽

Factor H ◽

Surface Proteins ◽

Host Immune System ◽

Dot Blot ◽

Binding Regions ◽

Far Western Blot

Abstract Streptococcus suis, a major emerging pathogen in swine and humans, expresses immunoglobulin G (IgG)-binding proteins (IBPs), which contribute to the ability of organism to evasion of host immune system. The objective of this study was to identify novel pig IgG (pIgG) and human IgG (hIgG)-binding proteins and characterize the binding regions of enolase from Streptococcus suis serotype 2 (S. suis 2). Here, four pIgG-binding proteins (pIBPs) and five hIgG-binding proteins (hIBPs) were identified from S. suis 2 surface proteins by 2D-Far-western blot assays. All the newly captured proteins were expressed and further confirmed their binding activity to pIgG or hIgG by Far-western blot and dot blot. In addition to previously identified factor H, fibronectin, collagen, fibrinogen, plasminogen and laminin, we also found that both pIgG and hIgG can specifically interact with enolase. Binding assays indicated that interactions of S. suis 2 enolase with pIgG and hIgG is primarily mediated by the enolase C-terminal portion (Enolase-C, a.a. 142-432). We found that hIgG exhibited stronger binding ability to Enolase-C than pIgG. Further analysis of the C-terminal regions of enolase (Enolase-C1 and Enolase-C2) suggested that the C-terminus possessed two different binding domains with distinct host IgG proteins. Strikingly, we confirmed that pIgG interacted with the Enolase-C1 (a.a. 142-271) and hIgG interacted with the Enolase-C2 (a.a. 271-432). These observations of enolase provide interesting insights in the pathogenesis of S. suis infection.

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Clinical Application of a Dot Blot Test for Diagnosis of Enteric Fever Due to Salmonella entericaSerovar Typhi in Patients with Typhoid Fever from Colombia and Peru

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.7.2.312-313.2000 ◽

2000 ◽

Vol 7 (2) ◽

pp. 312-313 ◽

Cited By ~ 4

Author(s):

Nora Cardona-Castro ◽

Eduardo Gotuzzo ◽

Monica Rodriguez ◽

Humberto Guerra

Keyword(s):

Typhoid Fever ◽

Clinical Application ◽

Sensitivity And Specificity ◽

Immunoglobulin G ◽

Good Predictor ◽

Enteric Fever ◽

Field Studies ◽

Dot Blot ◽

Laboratory Facilities ◽

Flagellar Antigen

ABSTRACT Clinical application of a dot blot test to detect immunoglobulin G (IgG) (88% sensitivity and specificity) and IgM (12.1% sensitivity and 97% specificity) against flagellar antigen from Salmonella enterica serovar Typhi was performed in Peruvian and Colombian patients with typhoid fever. This test can be used as a good predictor of serovar Typhi infection in regions lacking laboratory facilities and in field studies.

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Use of Protein a Conjugated to Peroxidase to Localize Immunoglobulin G in SSPE Ferret Brain

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100054376 ◽

1982 ◽

Vol 40 ◽

pp. 350-351

Author(s):

Hannah R. Brown ◽

Anthony F. Nostro ◽

Halldor Thormar

Keyword(s):

Immunoglobulin G ◽

Human Disease ◽

Measles Virus ◽

Subacute Sclerosing Panencephalitis ◽

Protein A ◽

Inflammatory Lesions ◽

Sspe Virus ◽

Specific Immunoglobulin ◽

Antibody Titers ◽

The Brain

Subacute sclerosing panencephalitis (SSPE) is a slowly progressing disease of the CNS in children which is caused by measles virus. Ferrets immunized with measles virus prior to inoculation with the cell associated, syncytiogenic D.R. strain of SSPE virus exhibit characteristics very similar to the human disease. Measles virus nucleocapsids are present, high measles antibody titers are found in the sera and inflammatory lesions are prominent in the brains. Measles virus specific immunoglobulin G (IgG) is present in the brain,and IgG/ albumin ratios indicate that the antibodies are synthesized within the CNS.

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A combined ultrastructural and gene molecular research for the relationship between human uterine cervical carcinoma and human papillomavirus

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010015856x ◽

1990 ◽

Vol 48 (3) ◽

pp. 204-205

Author(s):

Kun Lee ◽

Jingyi Si ◽

Ricai Han ◽

Wei Zhang ◽

Bingbing Tan ◽

...

Keyword(s):

Cervical Cancer ◽

Human Papillomavirus ◽

Intraepithelial Neoplasia ◽

Hpv Infection ◽

Dot Blot ◽

Homologous Sequences ◽

Normal Cervical Epithelium ◽

The Relationship ◽

Chronic Cervicitis

There are more supports for the view that human papillomavirus (HPV) infection might be an etiological factor in the development of cervical cancer when the association of persistent condylomata is considered. Biopsies from 318 cases with squamous cell carcinoma of uterine cervix, 48 with cervical and vulvar condylomata, 14 with cervical intraepithelial neoplasia (CIN), 34 with chronic cervicitis and 24 normal cervical epithelium were collected from 5 geographic regions of China with different cervical cancer mortalities. All specimens were prepared for Dot blot, Southern blot and in situ DNA-DNA hybridizations by using HPV-11, 16, 18 DNA labelled with 32P and 3H as probes to detect viral homologous sequences in samples. Among them, 32 cases with cervical cancer, 27 with condyloma and 10 normal cervical epitheliums were randomly chosen for comparative EM observation. The results showed that: 1), 192 out of 318 (60.4%) cases of cervical cancer were positive for HPV-16 DNA probe (Table I)

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