Monoclonal & Polyclonal Antibodies

Question 1

Antibodies specific for 1 antigen; made in laboratory; Ab in serum (by collecting blood); induce immune response from mice; risk disease in blood which contaminates serum

Answer 1

Monoclonal antibodies (mAbs)

Question 2

What is the 1st step in mAb production?

Answer 2

Choose an antigen especially responsible for pathogen attachment.

Question 3

What species is commonly used in mAb production and why?

Answer 3

Mice—easy to handle and harvest spleens from

Question 4

What does splenomegaly indicate after antigen injection? (Enlarged spleen due to immune response)

Answer 4

Immune response due to antigen-antibody reactions

Question 5

White blood cells from the spleen, including B cells

Answer 5

Splenocytes

Question 6

How are splenocytes isolated?

Answer 6

Spleen is crushed, filtered, diluted, lysed with RBC lysis buffer; RBCs removed, WBCs retained

Question 7

Why can’t plasma cells be cultured long-term?

Answer 7

not immortal and die after a short time

Question 8

Hybrid cells that are end product of fusion of Myeloma + Plasma cells

Answer 8

Hybridoma

Question 9

How are immortal plasma cells created?

Answer 9

Fuse plasma cells with myeloma (cancer) cells to form hybridomas

Question 10

Culture medium used for mAb production allowing only hybrid cells to grow; ensures only hybridomas grow—unfused cells cannot survive

Answer 10

Selective medium

Question 11

How is growth detected in selective medium?

Answer 11

red/pink to yellow indicates acidic pH due to cell metabolism

Question 12

How are monoclonal clones separated for mass production?

Answer 12

Transferred into 96-well plates for single-cell cloning

Question 13

What are two methods used to test specificity of mAbs? (Techniques used to detect antigen-antibody interaction)

Answer 13

ELISA and Western Blot

Question 14

Test specificity of mAbs whose specific binding causes color change (yellow) based on antibody presence

Answer 14

ELISA

Question 15

What happens in a non-specific ELISA test? (When antibody fails to bind to target antigen)

Answer 15

No color change after washing; antibody and substrate are removed

Question 16

Test specificity of mAbs which detects specific protein-antibody interaction using visible bands after transfer to PVDF membrane

Answer 16

Western Blot

Question 17

Gel electrophoresis method that separates proteins by size in Western Blot; proteins are separated and then transferred for detection using antibodies

Answer 17

SDS-PAGE

Question 18

mAb used in dogs with allergic dermatitis; blocks IL-31 to reduce itching

Answer 18

Cytopoint

Question 19

Monoclonal antibody used to treat osteoarthritis pain in cats; blocks Nerve Growth Factor (NGF)

Answer 19

Frune Vet mAb

Question 20

Monoclonal antibody for managing pain in dogs, also targets NGF to relieve osteoarthritis pain

Answer 20

Bedin Vet mAb

Question 21

Monoclonal antibody against a canine viral disease; blocks attachment of Canine Parvovirus, preventing viral entry

Answer 21

CPmA

Question 22

Antibodies that recognize multiple epitopes on a single antigen; Multiple antibodies produced by different B cells, recognizing different parts of the antigen

Answer 22

Polyclonal antibodies

Question 23

Ability to stimulate a strong immune response?

Answer 23

Immunogenicity

Question 24

Why should vaccines have high immunogenicity?

Answer 24

To ensure a robust and protective immune response

Question 25

1st step in pAb production?

Answer 25

Triggering immune response by introducing antigen: Inject antigen into rabbits (sometimes goats, pigs, or hamsters)

Question 26

What happens after antigen injection in pAb production?

Answer 26

B cells are activated and differentiate into plasma cells

Question 27

How are pAbs collected?

Answer 27

Antiserum is collected from blood and used directly

Question 28

What are the two techniques to test pAb specificity? (Lab assays that detect antigen-antibody binding)

Answer 28

ELISA and Western Blot

Question 29

Why might Western Blot be unreliable for pAbs?

Answer 29

May show multiple bands—false positives due to cross-reactivity or contaminants

Question 30

Key disadvantage of using pAbs in therapy?

Answer 30

Antiserum may contain pathogens or cause allergic reactions; not considered safe for therapy

Question 31

Residual fluid when blood forms clot where antibodies can be found

Answer 31

Serum