4. Lecture

Question 1

function of surface proteins: (capsid, envelope)

Answer 1

» shape, antigenicity, adsorption, HA activity, enzymes

for penetration or release of virions

Question 2

function of core proteins:

Answer 2

» NA protection, stabilization, enzymes for replication

Question 3

non structural proteins

Answer 3

coded in the viral genome
– not integrated in the virion
– present only in the vegetative virus
– early, immediate-early proteins
» regulation of the cell, inhibition of cellular defence
» enzymes for replication
– late proteins
» enzymes for structural protein maturation, virus
assembly
» (the majority of the late proteins are structural!)

Question 4

PAGE

Answer 4

protein investigation: SDS-Polyacrylamide-gel electrophoresis (PAGE)

separation of charged molecules in mixtures by their molecular masses in an electric field: number and size

linearization of the polypeptides to see bends

Question 5

immunoblotting aka. western blot

Answer 5

Transfer of viral proteins from PAGE into nitrocellulose filter

Question 6

immunoperoxidase staining used in

Answer 6

SDS-PAGE, Western blot, or in histology sections

- detection of antigen

Question 7

detection of antigens using immunoperoxidase staining

Answer 7

cover with primary Ab (specific to the Ag)
• incubation, washing
• cover with conjugate
– peroxidase labeled Ab
– specific to the primary Ab
• incubation, washing
• cover with substrate
• → color reaction

Question 8

MAb = monoclonal antibody production

Answer 8

Prod by single clone of B lymphocytes

Specific for a particular epitope

Question 9

how are lymphocytes isolated for monoclonal antibody production, and how does the production occur

Answer 9

in the spleen
B cells; Ab production ut no mitotic activity. the B cells are mixed with myeloma cells, which will fuse with B lymphocytes –> tetra-ploid hybridoma cells - which can multiply and produce antibodies

Question 10

testing the clones ( tetra-ploid hybridoma cells )

Answer 10

Testing the clones: ELISA, IF, IPO
–> epitope specific MAb’s
Large scale production can be done in the abdomen of mice

Question 11

how are viral lipids aquired

Answer 11

30-35% of envelope
made from cellular membrane structures (budding)
– phospholipid, cholesterol
– + virus specific proteins, glycoproteins
-> poxviruses, African swine fever virus: virus specific lipids too

Question 12

viral carbs types, found where

Answer 12

Ribose, deoxyribose
• in the nucleic acid
– Glycoproteins
• on the surface
• pox: in the core too