Ch 1:

GVHD: NGUYN TH ANH TH

NHM 1
THNH VIN NHM
- H Th Hoa
- Nguyn Th Xun Thy
- Phm Th Thoa
- Nguyn Th Xun Thy
- Nguyn Th Bch Lin
- Trn n
Khi nim
Cu trc
Phng
php sn
xut
u nhc
im
ng dng
Ni
dung
chnh
I) Khi nim
- Nanobody l mnh v nh nht ca khng th thiu
chui nh nhng c y chc nng v kh nng
lin kt vi khng nguyn nh mt khng th thng
thng.



II) Cu trc
- Nanobody l mt protein n gin
- Kch thc nh ch c 15 kDa ; vi 2,5 nm ng
knh v 4 nm chiu cao)
- Cha khong 120 axit amin
- Tng ng vi vng VH ca con ngi n 90%
-Vng CDR1 v CDR3 ko di vi bin i ln hn
v linh ng hn vng VH ca khng th thng
+ Vng CDR3 ca VHH cha 16-18 axit amin( ngi
v chut l 9-12 axit amin)

video
II) Cu trc
III) Phng php sn xut
1)T khng th chui nng

2) T khng th thng thng

III) Phng php sn xut
- T khng th chui nng
1. Immunize camelid with immunogenes
2. Collect blood from camelid
3. Isolate lymphocytes
4. mRNA isolation and reverse transcription polymerase
chain reaction (RT-PCR)
5. Generation of VHH library ~ 10
6
-10
7
VHH
clones are obtained
6. Specific VHHs are selected by panning
III) Phng php sn xut
IV) u nhc im ca Nanobody

1) u im

- Kch thc nh.

- C th nhn din
c cc epitope
thng hay n


IV) u nhc im ca Nanobody
1) u im

- D sn xut

- Tnh n nh cao

- Ha tan tt

IV) u nhc im ca Nanobody
2) Nhc im
- D b o thi: nanobody c kch thc 15kDa nh hn
nhiu so vi b lc thn( 60kDa) nn chng d dng i
qua b lc thn v b o thi ra khi c th.
Khc phc: tng hp cc VHH c th lin kt vi protein
albumin ln hn trong huyt thanh hay tng trng lng
phn t bng cch b sung cc nhm glycolpolyethylene
- S an ton v tim nng trong iu tr bnh ngi vn
cha chc chn: hin nay nanobody vn cn ang trong
qu trnh nghin cu v cha c ng dng rng ri.
Bng so snh
Khng th bnh
thng
Nanobody

c im di truyn
-C tt c cc ng
vt
-c m ha bi nhiu
gen

-Ch c lc v c sn
-c m ha bi mt gen duy nht

Cu trc
-Kch thc ln
(150kDa )
-C 2 chui nng v 2
chui nh


-Kch thc nh( 15kDa )
-Ch c 2 chui nng
-Vng CDR1 v CDR3 di hn
u im

- t b o thi -Xm nhp nhanh v su vo m
-C th nhn din c cc epitop
thng hay n
-Tnh n nh cao
-D sn xut
Nhc im
-Khng nhn din c
cc epitop n
-D b phn hy bi pH,
protease
-Tnh n nh thp
- D b o thi
V) ng dng
1) Nanobody c gi tr to ln trong xt nghim v
chn on bnh:



+ Vim d dy-rut:


V) ng dng
1) Nanobody c gi tr to ln trong xt nghim v
chn on bnh:


+ Bnh s va
ng mch:

V) ng dng
1) Nanobody c gi tr to ln trong xt nghim, chn
on v iu tr bnh:
+ Bnh ung th tuyn tin lit: nanobody c th phn bit
cc ng dng khc nhau ca PSA( mt loi khng
nguyn gy bnh ung th tuyn tin lit)
trong dng mu.
+ Bnh ng: nanobody gn vo mt protein o
trypanosome v hp nht vi enzyme apolipoprotein,dn
n ly gii trypanosome.
+ Bnh vim khp dng thp v bnh Crohn: nanobody lin
kt vi yu t gy hoi t khi u cu.

V) ng dng
2)Nanobody cng c th c s dng nhiu trong cng
ngh sinh hc ng dng.
+Protein dung hp ( chromobodies )