Point of care diagnostics- Lateral flow technology

Question 1

in lateral flow assay for the diagnosis of IPA which antibody is used

Answer 1

JF5 Ab

Question 2

J5F Ab is labeled with

Answer 2

colloidal gold- shows up red

Question 3

when antigen is present (person with IPA)

Answer 3

antigens bind to the labeled J5F Ab (comes before test and control line).

• these labeled J5F Ab flow up the pad via capillary action onto the membrane and binds to the immobile J5Ab
• unbound labeled antibodies will bind to the control antibodies (double line)

Question 4

when antigen is not present

Answer 4

labeled J5F Ab bind to the control immobilised antibodies alone (single line)

Question 5

outline protocol for how sample is prepared to be used in LFA

Answer 5

serum or haemorrhage BAlL combined with sample buffer and heated to 100 degrees

• centrifuged
• liquid at top added to LFA

Question 6

Lateral flow assay for IPA detects what

Answer 6

J5F antigen in human sera

Question 7

bronchoalveolar lavage used to

Answer 7

isolate aspergillus fumigateurs antigens

Question 8

how does BAL occur

Answer 8

bronchoscope put down trachea and into the left primary bronchus- water flushed and then sucked back up

Question 9

Non-invasive molecular imaging is

Answer 9

antibody guided

Question 10

antibody guided molecular imaging uses which machinery

Answer 10

ImmunoPET/MRI

Question 11

lung diseases

Answer 11

all look quite similar on CT scan

Question 12

on an x ray

Answer 12

• On an x-ray typically see diffuse lung patterning

Question 13

radiologists look for

Answer 13

abnormalities in the lungs

Question 14

radiological tests are very

Answer 14

• Radiologists looks for abnormalities in the lungs
• Very non-specific for fungal infections
• E.g. cant tell you directly what it is- can just tell that something is not right

Question 15

antibodies can be used to

Answer 15

show the presence of IPA in the lungs- very specific

Question 16

antibody has to be

Answer 16

humanised to prevent HAMA reaction

Question 17

immunoPET/MRI

Answer 17

immune positron emission tomography’ magnetic resonance imaging

Question 18

how to use immunoPET/MRI to identify IPA in mice

Answer 18

• knock out mice immunity
• 24h later challenge mice with spore of IPA (into trachea)
• inject tracer in tail
• in vivo and ex vivo bio distribution of 64-Cu- yelled antibody tracers

Question 19

why is immunoPET/MRI good

Answer 19

• non-invasive
• very specific
• no uptake into lungs of mice with other infections

Question 20

which antibody used

Answer 20

humanised JF5

Question 21

what causes mice to become immunocompromised

Answer 21

depletion neutrophil granulocytes in C57BL/6 mice via injection of anti-G3-1 antibody RB6-8C5

Question 22

how is humanisation of JF%

Answer 22

CDR grafting

Question 23

humanisation reduces the

Answer 23

immunogenicity of exogenic mAbs- improves activation of human immune system activation (ADCC/CMC)

Question 24

humanised antibodies

Answer 24

constant region is human

- variable light and heavy chains are grafter CDRs- e.g. complementary to JF5 CDR

Question 25

LFD using

Answer 25

mAb JF5 is a simple and quick front-line test

Question 26

immunoPET/MRI allows..

Answer 26

non-invasive detection fo aspergillus lung infection in vivo

Question 27

humanised version of mAb JF5 has entered

Answer 27

GMP production for first-in-human clinical trials 2017/18