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Laboratory of immunochemistry of glycoconjugates and Laboratory of lignin, Institute of Chemistry, SAS

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Title: Laboratory of immunochemistry of glycoconjugates and Laboratory of lignin, Institute of Chemistry, SAS


1
Laboratory of immunochemistry of
glycoconjugatesandLaboratory of
lignin,Institute of Chemistry, SAS
2
Preparation and immunological characterization of
glycoconjugates based on cell-surface
polysaccharide antigens from the yeasts of
Candida genus potential vaccines
Yeasts of Candida genus represent serious social
and medical problem. Our laboratory belongs to
those few departments in the world where the
potential vaccines against diseases caused by the
Candida yeasts are being developed.Our vaccine
constructs are based on polysaccharide from the
cell of yhe yeasts, which are responsible for the
antigenicity of the yeasts.Children lt 2 yrs,
elderly people and immunocompromised individuals
can produce sufficient level of IgG antibodies
only against T-cell dependent antigens
(polysaccharides are T-cell independent).
Conversion of polysaccharides to T-cell
dependence antigens can be achieved via
conjugation with protein carrier.
3
General strategy of glycoconjugate vaccine
preparationYeast cultivation (most frequently
present human pathogensC. albicans, C.
tropicalis, C. parapsilosis, C. glabrata),
isolation and purification of mannansMannan
derivatizationConjugation with
proteinImmunization of animal modelImmunological
characterization of the conjugate activity
4
Study of the structure and biological activity of
fungal polysaccharides at the Laboratory of
microbial polymers
  • By means of acidic and alkaline extractions,
    cellular ß-D-glucan was isolated from the cell
    walls of yeasts Saccharomyces cerevisiae and
    Candida albicans.
  • Using 13C-NMR spectroscopy and by observing the
    complex formation with Congo Red dye, it has been
    proven that ß-D-glucans form helical structures
    depending on the pH value (single and triple
    helices).
  • Water-soluble derivatives of S. cerevisiae glucan
    carboxymethyl- and sulfoethyl glucans were
    prepared by a chemical modification, and by means
    of the ultrasonic treatment, fraction
    precipitation, and liquid chromatography, their
    defined fractions with different degrees of
    derivatization and molecular weight values were
    obtained.
  • Water-soluble derivatives of S. cerevisiae glucan
    exterted protective effect in mice with
    experimental infection caused by bacteria
    Klebsiella pneumoniae.
  • Carboxymethyl glucan inhibited lipid peroxidation
    in liposomes and revealed antioxidant activity
    comparable to that of the established
    antioxidants such as D-mannitol, a-tocopherol,
    and ascorbic acid.
  • Carboxymethyl glucan and sulfoethyl glucan
    prevented growth and metastasis of Lewis lung
    carcinoma and lymphosarcoma tumors and showed
    synergistic antitumor effect with a known
    cytostatic drug cyclophosphamide, and moreover
    suppressed its toxic side-effects.
  • Water-soluble derivatives of S. cerevisiae glucan
    demonstrated antimutagenic and antigenotoxic
    effect and inhibited the oxidative degradation of
    DNA in lung cells.
  • The ability of carboxymethyl glucan to scavenge
    free radicals was for the first time directly
    proven by a spin-trap EPR spectroscopy.
  • Water-soluble derivatives of S. cerevisiae glucan
    revealed protective anti-arthritic activity in an
    experimental model of adjuvant arthritis in mice.
  • Antioxidant and antimutagenic properties were
    manifested also by other fungal polysaccharides
    glucomannan from the yeast Candida utilis and
    glucan-chitin complex from the filamentous fungus
    Aspergillus niger.

5
Synthesis and characterization of
neoglycoconjugates based on Vibrio Cholerae O1
LPS. Derivatization of LPS carboxylic groups
using different linkers and conjugation with
proteins
- disease caused by Gram negative bacteria Vibrio
Cholerae, every year over 100 000 dead -
prevention strict sanitation and vaccination -
cell surface of bacteria contains
lipopolysaccharides (LPS). Immunity against these
LPS prevents against disease.
- isolation of antigen (extraction of cell mass
and LPS purification), separation or destruction
of lipidic part Lipid A (whole LPS is responsible
for septic shock) - conjugate synthesis
(conversion to T-cell dependent antigen).
Conjugate consists of antigen covalently bound to
protein carrier (different geometry, valency
etc.) - conjugate characterization and in vivo
testing (animal model)
6
Lignin belongs among main biomass components -
chemical treatment of wood worldwide yields about
50x106 t/ year.
In Lignin Laboratory under the supervising of
professor Ing. B. Košíková DrSc. novel methods
were developed for use of lignin biomass
component
1. application of lignin in polymer blends 2.
preparation of novel lignin-based antimutagenic
and anticarcinogenic compounds
  • isolation and modification of various types of
    lignins
  • characterisation of rheological and
    physico-mechanical properties of
    lignin-polyolefin blends
  • examination of the stability of polyolefin
    blends during processing and life service
  • influence of lignin concentration on
    vulcanization, rheological and mechanical
    properties of rubber composites
  • determination of antioxidative effect of lignin
    samples on rubber blends

7
Reduction of oxidative damage of DNA by lignin
preparations
was prooved on - cells on hamster cells V79 -
human cells VH10 - human carcinoma cells Caco-2 -
primary testicular cells and lymphocytes isolated
from the rats in vitro - lymphocytes and
testicular cells isolated from the rats fed by
diet containing lignin ex vivo
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