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Production of monoclonal antibodies (mAbs) against oralcarcinoma & its bioconjugation with solid lipid nanoparticles (SLN) for drug delivery.

This preliminary study explores a dual strategy of augmenting anticancer therapy with Carboplatin-loaded solid lipid nanoparticles (SLNs) alongside antigen-specific antibodies through immunization of KB oral carcinoma cells. KB and Sp2/01 myeloma cells were cultured and assessed for viability. Female Balb/c mice were immunized with KB cells, resulting in significant weight gainand spleen enlargement, suggesting immune activation. Enzyme-linked immunosorbent assay (ELISA) confirmed the production of antigen-specific antibodies, as indicated by an increase in absorbance at a 1:1000 serum dilutio

Expression and characterization of a recombinant broadly-reactive monoclonal antibody against group 1 and 2 influenza viruses.

Production of broadly-reactive antibodies is critical for universal immunodiagnosis of rapidly-evolving influenza viruses. Most monoclonal antibodies (mAbs) are generated in mice using the hybridoma technology which involves labor- and time-consuming screening and low yield issues. In this study, a recombinant antibody based on a broadly-reactive mAb against the hemagglutinin (HA) stalk of H7N9 avian influenza virus was expressed in CHO cells and its biological characteristics, cross-reactivity and epitope recognition were identified. The variable genes of the parental antibody were amplified

[Preparation of monoclonal antibodies against enterovirus type 71 with an epitope-incorporated adenovirus type 3 vector].

Objective To develop the monoclonal antibodies (mAbs) against enterovirus type 71 (EV71). Methods Two neutralization epitopes, SP70 and SP55, from EV71 were cloned into the hexon gene of adenovirus type 3 to generate a recombinant adenovirus type 3 (R1R2A3) presenting SP70 and SP55 antigens. BALB/c mice were immunized with the R1R2A3. The mAbs were developed with hybridoma technology and were analyzed with microneutralizing assay, indirect ELISA, Western blotting and direct immunofluorescence assay (DFA). Results The study obtained four hybridoma cell clones, 2C4, D2C9, I2G2 and I12C3. ELISA s

Leishmanin skin test in guinea pig with a single purified protein of Leishmania major.

A series of hybridomas was produced by fusion of SP2/0 myeloma cells with spleen cells of mice immunized with Leishmania major (L. major). The reactivity of secreted monoclonal antibodies (mAbs) was evaluated against available leishmanin antigen by enzyme linked immunosorbent assay. Only one hybridoma designated as 7F9 secreted IgG1 mAb which was shown to be reactive with leishmanin. This mAb was further tested against four species of Leishmania (L. donovani, L. tropica, L. infantum, L. major) and a recombinant gp63. Among the four species tested it was shown to be only reactive with promastig…

Isolation and epitope characterization of human monoclonal antibodies to hepatitis C virus core antigen.

In this study we describe the establishment of two hybridoma cell lines secreting human monoclonal antibodies to the 22-kD nucleocapsid protein (core, p22) of the hepatitis C virus (HCV). For this purpose we isolated B lymphocytes from an anti-HCV positive blood donor and infected them with Epstein-Barr (EBV). We obtained several lymphoblastoid cell clones secreting antibodies to the recombinant HCV core protein. The B-cell cultures were oligoclonally expanded and two of them were fused with the (mouse:human) heteromyeloma cell line K6H6/B5. The resulting stable hybridomas produce antibodies o…

Localization of biologic functions of toxic shock syndrome toxin-1 by use of monoclonal antibodies and cyanogen bromide-generated toxin fragments.

Monoclonal antibodies (mAb) against toxic shock syndrome toxin-1 (TSST-1) were generated that block two of the most important biologic activities of TSST-1, nonspecific T lymphocyte mitogenicity and the suppression of immunoglobulin synthesis. Fourteen hybridomas producing antibody against TSST-1 were isolated independently. The culture supernatant and ascitic fluids from each were analyzed to determine the mAb isotypes. Seven of the mAb were IgG1, and the remaining seven were IgM; all the mAb had kappa light chains. Immunoglobulin was partially purified from hybridoma-generated ascitic fluid …

Activation of human monocyte cytotoxicity by natural and recombinant immune interferon.

Human T cell hybridomas were established by fusion of SH9 cells, the 6-thioguanine-resistant mutant line of human T lymphoma Hut 102-B2, with concanavalin A-stimulated human peripheral blood lymphocytes. Hybridoma line L38 produced a macrophage activating factor (MAF) with the ability to activate human peripheral blood monocytes to show enhanced cytotoxicity against human colon adenocarcinoma HT-29 cells in a 72-hr 125iododeoxyuridine-release assay. The L38 line was then cloned by the limiting dilution technique and two sublines, L38B and L38D, were found to produce high levels of MAF constitu…

A RAST assay for monitoring specific IgE antibodies in the mouse.

Mouse monoclonal anti-ovalbumin IgE was isolated from serum of BALB/c mice implanted with the IgE secreting hybridoma (14-205). Purified IgE was used to immunize rabbits and for the preparation of an antiserum specific for mouse IgE. The latter was used to develop a RAST assay to replace the PCA technique in monitoring antigen-specific IgE response in the mouse.

Preparation and Antigenic Site Identification of Monoclonal Antibodies against PB1 Protein of H9N2 Subtype AIV.

Recently, low pathogenic avian influenza virus (LPAIV), including H9N2 subtype, has been common clinical epidemic strains, and is widely distributed globally. The PB1 protein is a key component of the viral RNA polymerase complex (vRNP), and is vital to viral transcription and translation. In this study, to investigate the antigenic determinants in the PB1 protein, the truncated PB1 sequence (1bp-735bp) from H9N2 subtype AIV was amplified with PCR, and expressed in plasmid pET-28a (+). After purification, the recombinant PB1 protein was used to immunize BALB/c mice. Following immunization, hyb

[Preparation and identification of neutralizing antibodies to envelope protein domain III(ED III) of the four dengue virus serotypes].

Objective To prepare neutralizing monoclonal antibodies (mAbs) against envelope protein domain III of the four dengue virus serotypes (DENV-EDIII) and identify its specificity. Methods BALB/c mice were immunized with recombinant EDIII protein (rDENV-EDIII) of the four DENV serotypes. Hybridoma cells secreting DENV-EDIII antibodies were screened by indirect ELISA. The specificity of positive hybridoma cells were further tested by indirect immunofluorescence assay (IFA). The neutralizing activities of DENV-EDIII mAbs in vitro were determined by the enzyme-linked immunospot microneutralization te

[Preparation of tiamulin artificial complete antigen and development of its monoclonal antibody].

Objective To develop high-sensitivity and high-specificity monoclonal antibody against tiamulin (TML). Methods Using oximation and carbodiimide method, TML was conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) to prepare the immunogen TML-BSA and coating antigen TML-OVA, respectively. BALB/c mice were immunized with TML-BSA. The anti-TML monoclonal antibody hybridoma cell lines were screened by indirect ELISA after cell fusion. The ascites were prepared by in vivo induction method. The monoclonal antibody was purified from ascites by caprylic acid-ammonium sulfate method and identif

Tracking the in vivo localization of streptococcal cell membrane (SCM) monoclonal antibodies: potential model for post-streptococcal sequelae.

Placement of ten different anti-streptococcal monoclonal antibody (mAb) secreting hybridoma cells, as well as positive and negative controls into animals and sacrificing on a daily basis showed a difference in the tissue sites of accumulating mAb as noted by fluorescent antibody testing. Initial binding of the mAb was noted by day two on the kidney GBM in all the animals. Striated muscle tissues tested positive starting at day nine with only four of the mAbs, by which time the GBM was strongly positive. Fluorescent antibody testing of heart and skeletal muscle from animals in which one of the

Antiphospholipid antibodies are associated with enhanced oxidative stress, decreased plasma nitric oxide and paraoxonase activity in an experimental mouse model.

OBJECTIVE: Oxidative stress contributes to atherosclerosis, and evidence of enhanced oxidative stress exists in antiphospholipid syndrome (APS). In a non-lupus murine model, we evaluated whether anticardiolipin (aCL) antibodies could affect the oxidant/antioxidant balance as an early biochemical step of APS. METHODS: Hybridomas producing human and murine aCL and anti-beta(2)-glycoprotein I (abeta2-GPI) monoclonal antibodies were injected into three groups of five female BALB/c severe combined immunodeficiency (SCID) mice. Corresponding hybridomas secreting non-antiphospholipid antibodies of t…

Low molecular weight IgM and CD5 B lymphocytes in rheumatoid arthritis.

OBJECTIVES: To evaluate the role of low molecular weight (LMW) IgM and CD5 B cells in rheumatoid arthritis (RA) and to explore the possibility that LMW IgM is derived selectively from this subset of B cells. METHODS: LMW IgM in sera and culture supernatants was detected by a sensitive immunoblot technique with an enhanced chemiluminescence detection system. CD5 B cells were determined by FACScan cytometry. In vitro studies were established in culture plates containing pokeweed mitogen with or without 2-mercaptoethanol (2-ME). Supernatants were obtained from CD5 positive hybridomas and CD5 neg…

The CD4 transmembrane GGXXG and juxtamembrane (C/F)CV+C motifs mediate pMHCII-specific signaling independently of CD4-LCK interactions.

CD4+ T cell activation is driven by five-module receptor complexes. The T cell receptor (TCR) is the receptor module that binds composite su … under purifying selection (Lee et al., 2022). Expressing mutants of these motifs together in T cell hybridomas increased CD4-LCK association but reduced CD3ζ, ZAP70, and PLCγ1 phosphorylation levels, as well as IL-2 production, in response to agonist pMHCII. Because these mutants preferentially localized CD4-LCK pairs to non-raft membrane fractions, one expl …

[Identification of Vibrio cholerae serogroups O1 and O139 by monoclonal antibodies in the slide agglutination test].

The agglutinating properties of MCA-O1 of the IgG class and MCA-O139 of the IgM class towards epitopes of O-antigen of Vibrio cholerae O1 and accordingly Vibrio cholerae O139 were studied. The ascitic and cultural fluids by hybridomas F8G12 and D11 deposited in the specialized Collection of Cell Cultures of Vertebrates (Saint Petersburg) under RKKK (II) 386 D and RKKK (II) 674 D were the sources of monoclonal immunoglobulins. The advantage of diagnostic monoclonal immunoglobulins is that they are distinguished for strict specificity and their use in practical health care contributes to the hig…

Growth inhibition of human breast carcinoma and leukemia/lymphoma cell lines by recombinant interferon-beta 2.

Recombinant human interferon-beta 2 was produced in Escherichia coli by direct expression of cDNA encoding the mature protein sequence. At concentrations that stimulate DNA synthesis and growth in B-cell hybridomas and plasmacytomas, the cytokine was found to exert a strong inhibition on the growth of a number of carcinoma and leukemia/lymphoma cell lines. This antigrowth effect was observed in clonogenic assays and by measurements of cell number and thymidine incorporation in growing cultures. The effect was blocked by antibodies to a synthetic peptide from the N terminus of the molecule. Nor…

Моноклональные антитела для обезьяньего оспового вируса: подготовка и применение.

В результате двух экспериментов с синтезом клеток NSO миэломы было выявлено 30 линий гибридных клеток, которые постоянно и активно скрывали моноклональные антитела (MoAb) до вируса обезьяны оспы. Восемь строк, как представляется, производят антитела для конкретных антигенных детерминант. Была описана изотипная и серологическая реактивность MoAb до обезьяньего оспового вируса, и был определен кариотип нескольких гибридом. Из MoAb был подготовлен фермент с надписью "Супруга" с выборочной реакцией на вирус обезьяны. Этот брак позволил выявить вирус обезьяны в материалах пациентов. Она также помог

Совпадение в аннотации оригинала
Two fusion experiments with NSO myeloma cells yielded 30 hybrid cell lines which continuously and actively secreted monoclonal antibodies (M … and serologic reactivity of MoAb to monkey pox virus was characterized and the karyotype of several hybridomas determined. An enzyme labelled conjugate has been prepared from MoAb selectively reacting with monkey pox virus. This conjugate allowed the identification of monkey pox virus in the materials from patients. It also helped to determine that an orthop …

Identification in several human myeloid leukemias or cell lines of a DNA rearrangement next to the c-mos 3'-end.

A characteristic DNA rearrangement, the loss of an EcoRI cleavage site next to the 3'-end of the human c-mos gene, has been found to be freq … ained from the marrow of the same patients. This DNA rearrangement is maintained in three different hybridomas derived by fusion of CM-S cells with normal human embryo hepatocytes.

Antigen processing of the heptavalent pneumococcal conjugate vaccine carrier protein CRM(197) differs depending on the serotype of the attached polysaccharide.

The pneumococcal (Pn) conjugate vaccine includes seven different polysaccharides (PS) conjugated to CRM(197). Utilizing antigen-processing cells and a CRM(197)-specific mouse T-cell hybridoma, we found that the serotype of conjugated PnPS dramatically affected antigen processing of CRM(197). Unconjugated CRM(197) and serotype conjugates 14 and 18C were processed more efficiently.