CNG NGH

S N XU T
VACXIN
(Vaccine prodution technology)

I. KHI NIM V MN HC

Cng ngh sn xut vacxin (vaccine production

technology)l mt mn khoa hc nghin cu
cong ngh sn xut vc xin dng phng bnh
truyn nhim cho ng vt v con ngi v mt
s mc ch khc, nhm phc v i sng v
bo v sc khe ca con ngi.

Ni dung chnh ca mn hc bao gm:

1. Tm hiu v lch s v phn loi vacxin
2. Nghin cu cng ngh sn xut vacxin
3.Nghin cu cc quy trnh kim nghim vacxin
4.Nghin cu phng php s dng vacxin.
7. Ti liu hc tp
+ Gio trnh Min dch hc ng dng
Nguyn B Hin - Trn Th Lan Hng
Nh xut bn Nng nghip, H Ni
+ Gio trnh Min dch hc
Nguyn B Hin Trn Th Lan Hng
8. Ti liu tham kho
+ Min dch hc
V Triu An Jean Claude Homberg
Nh xut bn Y hc, 1997
+ Min dch D ng hc c s
V Minh Thc
Nh xut bn i hc Quc gia H Ni, 2005
+ Min dch hc
Nguyn Ngc Lanh, Vn nh Hoa
NXB Y hc, H Ni, 2006
+ Vi sinh vt -Bnh truyn nhim vt nui
Nguyn B Hin,Nguyn Quc Doanh, Phm S
Lng, Nguyn Th Kim Thnh, Chu nh Ti.
NXB Gio dc, H Ni, 2008
Chng I
KHI NIM V PHN LOI VACXIN
-
Mc tiu :
Nm c c tnh, thnh phn v cch phn loi vacxin
-
Kin thc trng tm :
Khi nim v nguyn l ca vacxin
c tnh c bn ca vacxin
Thnh phn ca vacxin
Proteosom, liposom v coclet
Phn loi vacxin
I. LCH S PHT HIN V DANH PHP

Vacxin c chc nng to MD cho c th ngi v ng

vt chng li cc bnh truyn nhim.

T xa xa, con ngi nhn thy c nhng bnh

truyn nhim ch gp mt s loi ng vt v trong
cng mt v dch c th c c th mc nng, c c th
mc nh.

Mt khc, c nhng bnh sau khi b bnh qua khi th

vnh vin khng b mc li, tc l con ngi bit ti
nhng g m ngy nay chng ta gi l MD.

i vi cc bnh truyn nhim nguy him, con ngi

c mt qu trnh u tranh phng chng ginh git ly
s sng.

Ngay t khi s sng xut hin, trong s u tranh

sinh tn ca cc loi sinh vt, bnh tt xut hin,
cc loi sinh vt tc ng ln nhau, gy ra qu
trnh bnh l.

c bit l s tc ng k sinh ca cc loi VSV v

KST ti cc ng vt bc cao, gy ra nhng bnh
tt him ngho, to ra nhng v dch thm khc v
cp i nhiu sinh mng.

ginh git s sng, con ngi tm mi bin

php nhm hn ch nhng tc ng c hi ca
cc i tng gy bnh.
Bnh di
U MA
Herpessimplex
HPV
AIDS
Quai b
zona
Bnh phong
Un vn ngi ln
Giang mai

phng chng bnh di, nh khoa hc ngi Hy Lp

(384 322 trc CN) ngh cht b t chc b ch
di cn.

Vi bnh u ma
Ly vy mn u ma phi kh, tn nh cho ngi lnh ht
gy bnh nh, to MD.
Phng php ny mang li hiu qu nht nh nhng nguy
him l khng kim sot c liu lng nn c th gy
cht ngi hoc to ra dch trong cng ng.

Nm 1798, Edward Jenner, mt thy thuc vng

Gloucestershive (Anh) dng dch m trong mn u b
chng cho ngi to MD.

y l pht minh quan trng trong s pht trin ca MD

hc,m u cho nghin cu v kh nng bo v c hiu
ca c th chng li cc tc nhn gy bnh.

Jenner:
(t 1796)
S dng cc virus
cng c ca ng
vt gy min dch cho
ngi.

Sau , L.Pasteur:
(t 1800)
Trn c s ny ch
to thnh cng cc
vcxin phng bnh Di,
nhit thn v t huyt
trng.

ghi nhn, trong hi ngh v danh php quc t,

ngi ta ngh gi tt c cc ch phm sinh hc
c nguyn l phng bnh nh vy vi mt tn
chung l vacxin xut pht t t vaccinia tn
ca virus u b.

Danh php gm 2 t ghp: t u: vacxin

t sau: tn bnh

Hin nay cc thut ng v cc sn phm sinh hc

dng trong th y thay i gia cc quc gia.

M:

V d:
O
Thut ng vacxin dng ch nhng sn phm
cha virus hoc nguyn trng sng hay nhc
c, vi khun sng hoc cc axit nucleic.
O
Cc sn phm cha vi khun cht v cc VSV
khc c gi tn nh bacterin, cht chit ca vi
khun (bacterial extract), cc cht di n v
(sub units), c t vi khun (toxoid).

Chu u:
O
Cc sn phm dng trong th y c gi l cc
sn phm s dng cho ng vt nhm to ra
MD ch ng hoc b ng hoc chn on
tnh trng MD.

Trong chng ny, thut ng vacxin bao gm

tt c cc sn phm to ra kch thch sinh MD
ch ng cho c th ng vt chng li bnh

Vic s dng ny ph hp vi thut ng quc t,

khng lin quan n:
- Cc sn phm sinh hc sinh MD th ng
- Cht kch thch MD
- Cht iu tr d ng hoc chn on bnh.

Vacxin c coi l thnh tu v i nht ca y hc

hin i.

Cng tc tim chng c thc hin tt c cc

quc gia v thc s tr thnh tm l chn
phng chng nhiu bnh truyn nhim ngi v
ng vt.

Tuy nhin vn cn nhiu bnh truyn nhim him

ngho do cc VSV, c bit l do virus gy ra vn
cha tm c vacxin d phng.

Hin nay, vic nghin cu ci to cc vacxin

hin c, ch to cc vacxin mi ang l mc tiu
phn u ca cc nh khoa hc.
Qu trnh pht sinh v ly truyn ca bnh
truyn nhim

Chut

Ngi

Ong


Rui



Ln b bnh


G b bnh



Vt b bnh




b bnh



n ln


n tru
b


n Vt


n g

2.1. Khi nim

Theo quan im trc y:

O
Vacxin l mt ch phm sinh hc trong cha chnh mm
bnh hoc khng nguyn ca mm bnh gy ra mt bnh
truyn nhim no cn phng (nu l mm bnh th phi
c git hoc lm nhc c bi cc yu t vt l, ha hc
v sinh vt hc).
O
Khi s dng cho ng vt, vacxin to ra mt p ng min
dch ch ng gip ng vt chng li c s xm nhim
ca mm bnh tng ng.

Cch hiu ny c hnh thnh trn c s thc t sn

xut vacxin
O
V d: vacxin nhit thn c lm t vi khun nhit thn
nhc c, vacxin phng lao c lm t vi khun lao bin
d (BCG), vacxin t huyt trng c lm t vi khun t
huyt trng c v hot, vacxin un vn c lm t
ngoi c t c gii c

Ngy nay, khi nim v vacxin c s thay i.

O
N khng ch cn l ch phn t vi sinh vt hoc
k sinh trng c dng phng bnh m cn
c lm t cc vt liu sinh hc khc (khng vi
sinh vt) v c dng vi mc ch khng phng
bnh.

V d: vacxin chng khi u lm t t bo sinh khi u,

vacxin chng th thai lm t receptor ca trng

Nhng d l vacxin c ch to t vt liu no v

c dng vi mc ch g th thnh phn buc phi
c vacxin l khng nguyn v khi a vo c th
ng, khng nguyn s gy ra p ng min dch.

Nh vy hin nay vacxin c hiu vi

khi nim rng hn:
O
Vacxin l ch phm sinh hc cha
khng nguyn c th to cho c th mt
p ng min dch v c dng vi
mc ch phng bnh hoc vi mc
ch khc.
1.3. Nguyn l

Vacxin to ra trong c th sng mt p ng MD

ch ng.

H thng MD ca c th hot ng, sinh ra KT c

hiu chng li nhng Epitop ca yu t gy bnh.

C th s dng vacxin xut hin trng thi MD thu

c ch ng nhn to c kh nng chng li s
xm nhim ca yu t gy bnh tng ng.
1.4. c tnh c b n c a m t vacxin
Vacxin phi c 4 c tnh c bn :
O Tnh sinh MD hay tnh mn cm
(immunogenicity)

L kh nng gy ra p ng MD dch th
hoc t bo hay c hai.

Tnh sinh MD ph thuc vo KN v c th

nhn kch thch.

C ngha l ph thuc vo tnh l ca KN,

ng a ca KN v c a ca mi c th
ng vt.
O. Tnh KN hay tnh sinh KT

Mt vacxin khi a vo c th phi c kh

nng kch thch c th sinh ra KT.

Yu t gy bnh c th c nhiu Epitop

khc nhau.

Trong c th c Epitop qu nh (Hapten)

khng c tnh sinh KT, mun chng sinh KT
cn i chng thnh c tnh KN, bng cch
kt hp chng vi mt protein mang ti v
hi.
. Tnh hiu lc

L kh nng bo h ng vt sau khi c

s dng vacxin.

Mt vacxin a vo c th, nhiu KT c

to ra nhng khng phi loi no cng c
hiu lc tc l tiu dit c yu t gy
bnh.

Do yu t gy bnh c nhiu Epitop khc

nhau nn trong bo ch vacxin trc tin
phi lm sao cho p ng MD chng li
nhng Epitop thit yu

Trong NC sn xut vacxin hin nay ngi

ta c gng phn lp nhng KN hay Epitop
thit yu lm cho vacxin c thun
khit v tin ti c th tng hp c
chng.
O
V d:

Vi virus Gumboro: protein VP2

Virus cm gia cm : H v N

Virus vim gan B : HBS ( KN b mt )

Hiu lc ca vacxin c nh gi qua thc

nghim nhng ch yu nh gi trn thc a sau
khi tim chng
Mc MD qun th, c th da vo lng KT
trung bnh trong HT v t l bo h trong qun th.

Trn VTN: nh gi mc p ng MD
sau tim chng vacxin v hiu lc bo h l
ng vt qua th thch cng c.
O
Th nghim thc a: Vacxin c tim chng
cho mt qun th V , nh gi kh nng bo h
khi ma dch ti ng thi th thch cng c
1 nhm ngu nhin.

Vacxin c hiu lc l gy c MD mc
cao v bo v c th ng vt lu bn.

Tuy nhin, hiu lc ca 1 vacxin ph thuc

vo nhiu yu t: bo qun, vn chuyn v k
thut tim phng.

V vy, ngi ta xy dng 1 mn khoa hc

mi gi l vacxin hc (vaccinology) mc
ch l nghin cu mi bin php t lc sn
xut n lc tiu dng tng tnh hiu lc
ca vacxin.
O Tnh an ton:
y l c tnh rt quan trng.

Sau sn xut vacxin phi c c quan kim nh

nh nc kim tra v: v trng, thun khit v v c.
O
V trng : Khng c nhim cc VSV khc.
O
Thun khit: Khng c ln cc thnh phn KN
khc c th gy ra cc phn ng ph.
O
V c : Liu s dng phi thp hn rt nhiu
so vi liu gy c.

Vacxin phi c th tnh an ton trong phng TN,

thc a, quy m nh v i tr.

Tn sut v mc nng nh ca cc phn ng ph

nu c phi c xc nh trc khi c em ra
dng chung, nhng vn phi c theo di cn thn.
1.5. Cc loi vacxin
+
C th chia vacxin lm cc loi sau:

Vacxin cht (v hot)

Vacxin sng

Vacxin di n v

Vacxin th h mi sn xut bng

cng ngh gen
1. Vacxin cht

L loi kinh in nht

Nguyn tc l lm cht yu t gy
bnh (virus hoc vi khun) nhng vn
gi c tnh mn cm v tnh khng
nguyn.

Vacxin loi ny ch yu gy p ng
MD kiu dch th.
+
Phng php lm cht yu t gy bnh
C 2 phng php
-
Phng php ha hc: Dng cc ha cht
. Nh formol git cht vi khun.
V d: vacxin t huyt trng ln v hot
. Vi virus c th dng cc cht kh c hot tnh
cao nh Ethylenamine, propiolacton. Nhng ha
cht ny v hot hon ton virus nhng khng lm
bin i protein cu trc.
V d: vacxin bi lit dng tim ngi
vacxin di bt hot dng propiolacton
vacxin LMLM dng Ethylenamine
-
Phng php vt l: sc nng, tia x (X, UV).
u, nhc im ca vacxin cht

u im: Khng c, khng gy nhim mi trng,

tnh an ton cao.

Nhc im:
-
Thi gian MD ngn do lng KN c nh v t dn
-
Liu lng tim ln do kh tim v d gy p xe.
-
MD xut hin chm, gy MD t bo km.
-
Khng can thip trc tip vo dch
-
Phi a vacxin nhiu ln, tng nguy c d ng.

Do l mm bnh cng c, nu bt hot khng tt s

c nguy c pht dch.
V d:
1 v dch bi lit M do s dng vacxin bi lit
v hot nhng khng trit .
1.5.2. Vacxin sng

L vacxin c sn xut t chng virus hay vi

khun cn sng,Khng cn tnh gy bnh cho
ng vt c tim phng
C kh nng gy p ng MD mnh, chng nhn
ln trong c th vt ch v to ra s kch thch
ca KN trong mt khong thi gian.

Vacxin sng bao gm:

Vacxin nguyn c

Vacxin v c

Vacxin nhc c
+
Vacxin nguyn c:

Dng chng virus nguyn c c quan h t loi

ng vt khc.
V d: Dng virus u b lm vacxin phng
bnh u ngi.

a vo c th virus c c lc hoc gim mt

phn c lc theo con ng thc nghim: c
lc ca virus s gim i khi chng c a vo
c th theo ng thc nghim (khng ging s
xm nhp ca chng trong t nhin).
V d: Tim phng hi chng vim phi
ngi bng adenovirus sng.

Vacxin v c (vacxin nhc c t nhin):

Vacxin c sn xut t nhng chng VSV
v c phn lp trong t nhin.

Vacxin nhc c ha:

c sn xut t nhng chng VSV sng c
c lc yu, khng c kh nng gy bnh
cho ng vt c tim chng.

Cc chng VSV ny c lm gim c lc

bng cc phng php: vt l, ha hc, sinh
vt hc v cng ngh gen.
Phng php lm gim c VSV
+
Gim c bng nhit :

VSV gy bnh thng nhy cm vi t

0
; Nu nui cy
chng nhit khng ph hp, VSV s gim c lc
nhng vn gi c tnh KN.

V d:
O
Vacxin nhit thn: nui VK 42 43
0
C/15- 20 ngy, VK
mt kh nng hnh thnh gip m, c lc gim, s dng
lm vacxin
O
Vacxin Sabin dng ung chng bi lit:
Chn cc chng virus bi lit t bin, cho nhn ln nhiu
ln trong t bo thn kh, nui cy T
0
thp.
Virus c th nhn ln trong tuyn nc bt ng tiu ha
nhng khng xm nhp c vo m thn kinh do khng
gy chng bi lit na.
+
Gim c bng yu t ha hc
O
V d:

Vacxin BCG (Bacterium Calmette Guerin)

L mt chng TK lao b M.T. bovinus

c c lc cao, nui cy trong mi trng c mt

b trong 13 nm

Sau 230 ln cy chuyn, vi khun khng cn

c, c s dng sn xut vacxin BCG.
+
Gim c bng phng php sinh vt hc
. Phng php gim c VSV c in
. Phn ln vacxin virus s dng cho ngi, ng
vt sn xut theo phng php ny.
. Ngi ta cy chuyn VSV nhiu i qua mi
trng t cm th
. VSV khng iu kin thc hin y chu k
sng, gy thay i h gen thay i c lc,
kh nng gy bnh.

V d:
O
Virus cng c DTL tim truyn lin tc 155
i qua th s thu c ging virus nhc c
DTL.
O
Virus dch t vt chuyn 41 46 i qua phi
g.
O
Virus vim gan vt typ I cy chuyn 54 i qua
phi g.
O
Virus bi lit c cy chuyn nhiu i qua
mi trng t bo thn kh.
O
Virus si c nui cy qua t bo x phi g.
+
u im ca vacxin sng:
-
To MD nhanh, mnh; MD tn ti lu bn
do VSV vn c kh nng nhn ln v tn ti
lu trong c th c tim chng.
- To min dch t bo cao hn vacxin cht.
- C th dng can thip trc tip vo dch.
-
Liu lng t, d tim chng.
+
Nhc im ca vacxin sng :
- Mc an ton thp do t bin dn n s
tr li cng c.
- Tp nhim virus trong nui cy t bo.
V d:
T bo thn kh c th tp nhim vi SV40
(Simianvirus)
- Kh bo qun, chi ph ln.
- M t s loa i khng s dng c cho ng
vt mang thai.
- Khng dng cho nhng vng an ton dch.
Bng so snh gia 2 loi vacxin
Loi vacxin
Ch tiu
Vacxin sng Vacxin cht
- Phng thc sn xut
- Tnh n nh
- Liu lng
- S ln a vacxin
- Cht b tr
- Thi gian min dch
- p ng MD t bo
- S tr li cng c
n gin
Tng i n nh
Thp
1 ln
Khng
Di
Tt
C th c
n gin
n nh
Cao
Nhiu ln
C
Ngn
Km
Khng
1.5.3. Vacxin di n v

L vacxin cha KN tng i tinh khit phn lp t

virus hay vi khun gy bnh

Mt s VK gy bnh bng c t: Cl.tetani,

Corynebacterium.diphtheria, ngi ta nui cy VK,
chit tch c t, gim c bng yu t ha hc hoc
vt l theo nguyn l ca vacxin cht. Cc c t mt
hot tnh c gi l gii c t (anatoxin) v c
dng lm vacxin.

Phy khun t gy bnh nh Enterotoxin, c t ny

gm 1 di n v A rt c v 5 di n v B khng
c, nhng B li c kh nng sinh khng th bo v
nn ngi ta nui vi khun, tinh lc Enterotoxin, tch
di n v B dng lm vacxin chng bnh t.
+
1 s bnh do virus gy ra, vacxin khng dng
ton b ht virus m ch dng 1 vi thnh phn
ca virus: protein capxit hoc glycoprotein v
ngoi do vacxin cn c gi l vacxin di
ht (subvirion).
+
Khng dng virus nguyn vn l do:
O
Nhiu loi virus khng c kh nng nhn ln cc
t bo nui cy (virus vim gan B).
O
Mt s virus c coi l rt nguy him nn khng
m bo an ton khi sn xut k c vacxin sng
hoc cht (HIV)
O
Mt s vacxin nu sn xut t virus nguyn vn c
th nh hng n c th do gy phn ng ph
(vacxin cm).

Lu : vic tng tinh khit ca vacxin

c th dn n mt tnh sinh MD hoc s b
cc enzym ph hy trc khi kch thch
min dch.
V vy loi vacxin ny i hi phi c vt
mang hay cht b tr, v d nh mui
nhm.

Vacxin di n v c mc thun nht

v tinh khit hn ton b VSV cho nn tnh
mn cm, tnh sinh KT v tnh hiu lc u
cao.
Cc protein capxit hoc glycoprotein v ngoi
thng gn vo receptor trn b mt mt s
loi t bo k ch, hoc thu c t huyt
tng bnh nhn, sau lm bt hot
(khng nguyn b mt ca virus vim gan
HBSAg) hoc c th sn xut bng k thut
ti t hp ADN...
Cc vacxin subvirion nh:
- vacxin HA chng virus cm typ A, B
- vacxin HBS chng virus vim gan B
1.5.4. Vacxin th h mi sn xut bng cng
ngh gen
Khi nim

Nhng tin b v KH - KT trong lnh vc VSV,

MD hc, sinh ha , c bit l k thut gen hc
v cng ngh sinh hc phn t m ra mt
hng ng dng mi l nghin cu sn
xut cc loi hnh vacxin bng cng ngh gen.

Nhng vacxin to ra bng phng php trn

gi l vacxin th h mi nhm phn bit vi
cc loi vacxin c c nghin cu sn
xut bng phng php cng ngh truyn
thng.

Mt vacxin c gi l vacxin th h mi
phi l thnh phm ca mt quy trnh c
s can thip, s dng, thao tc ca cng
ngh gen.

Hin nay nhiu loi vacxin th h mi

v ang c a vo s dng c hiu
qu, gp phn vo vic phng chng bnh
tt cho ngi v ng vt.
Nguyn l

Trong mt loi vacxin, yu t quyt nh

tnh sinh MD chnh l phn protein c
bit c trn b mt ca VSV gy bnh.
.Thnh phn protein ny c gi l KN
v do mt gen hay mt s gen c trong
h gen ca VSV gy bnh quyt nh
tng hp nn.
.Nhng gen chu trch nhim tng hp
protein KN c gi l gen KN.

Nu tch gen KN khi vt liu di truyn

ca VSV ri ghp vo mt h thng
plasmid vector thch ng no th gen
KN ny vn hot ng nh khi tn ti trong
h gen ca VSV ch v phn t protein KN
c tng hp ra vn c th c chc nng
nh c, tc l c tnh sinh min dch.

Ch phm protein KN c to ra bng

cng ngh gen nh th c gi l
vacxin th h mi vacxin cng ngh gen
Phn loi
Vacxin th h mi c nhiu loi. Cn c vo ngun KN nhn
ln c hay khng nhn ln trong c th ng vt, ngi ta
chia vacxin th h mi lm 2 nhm:
a. Vacxin c KN sng c nhn ln:
C Vacxin ti t hp c vector dn
truyn
C Vacxin axit nucleic (vacxin ADN)
C Vacxin xa gen c
b. Vacxin c KN khng nhn ln:
C Vacxin cha KN l protein sn xut
bng k thut gen
C Vacxin n c
C Vacxin peptit tng hp
a. Vacxin c KN sng c nhn ln, bao gm:
a. Vacxin c KN sng c nhn ln, bao gm:
C Vacxin ti t hp c vector dn truyn
Loi vacxin ny cha 2 thnh phn chnh:
- on ADN cha gen m ha cho KN chnh
c tch ra t VSV gy bnh.
-
H gen ca vector dn truyn.
Ngi ta tch ri gen KN t VSV gy bnh ri ghp vo
h gen ca vector dn truyn l plasmid hay VSV ri
a vo vt ch.
L SV sng nn khi gy nhim, chng s nhn ln do
ngun gen KN v sn phm ca gen KN l protein KN
lun c sn xut ra to MD lu bn cho c th.
+
Cc vector dn truyn:

Hin nay cc vector dn truyn c chn

thng l nhng sinh vt:
- Vi khun
- Virus
- Nm men
- Thc vt
Thng c th nhn ln c nhiu loi
ng vt v c lm gim c hoc v
c bng k thut gen.
Vi khun S. typhimurium:
L loi VK khng c, c chn lm vector
dn truyn v c cc u im:
- D s dng qua ng tiu ha.
- C th tn ti v nhn ln t chc Lympho
ng tiu ha, cung cp protein KN bn
vng gy p ng MD ton din: dch th,
t bo v min dch cc b.
- Vic nui cy VK ti t hp gen ny d thc
hin v thi gian sn xut rt ngn.

Hin nay S.typhimurium c s dng ph

bin lm vector dn truyn sn xut cc
vacxin: cm, vim gan B, st xut huyt,t,
lin cu khun, KST st rt...

Ngoi S. typhimurium, cn s dng vi khun

E.coli v mt s loi nm men nh:
- Pichia pastoris
- Saccharomyces cerevisiae


+ Virus u b ( vaccinia):
Thuc nhm Orthopoxvirus c s dng
mt thi k di lm vacxin phng bnh u
ma.

H gen ca virus u c 200.000 nucleotit

C kh nng tip nhn nhiu gen ngoi lai c
di 25.000 nucleotit l vector l tng
sn xut vacxin a gi

Mt khc virus u:
- C kh nng gy nhim vo nhiu loi vt
ch nhng khng gy bnh
- C th nhn ln trong nhiu loi t bo nui
cy, thm ch nhiu dng t bo m
chng khng hon thin c chu trnh nhn
ln khng b dung bo thun tin lm
vector.

Virus u ti t hp c th biu hin gen KN

sm v sn xut ra protein KN gy p ng
MD ton din.
S to vacxin ti t hp
vct dn truyn
Nui cy t
bo
chn lc
virus u
vect
virus u
ti t hp
gen
vacxin
khi pht
ATG
kt thc
TAA
ct
ni
gen
vacxin
im
mm
v tr
ct
vng
khi
ng
Plasmi
d
Plasmid
ti t
hp
Cc bc to vacxin ti t hp c vect dn
truyn:
- Ct on gen vacxin
- Gn on gen vacxin vo Plasmid
- Chuyn np cc Plasmid gn gen vacxin
vo
virus u virus u ti t hp.
Hin nay s dng virus u lm vect dn
truyn sn xut mt s vacxin ti t hp phng
bnh cho vt nui:
-Gi gen m ho protein F ca virus Newcastle
vo virus u to vacxin ti t hp phng
bnh Newcastle
-Gi gen m ho protein HN vo virus u to
vacxin ti t hp phng bnh cm gia cm.

Vacxin vim gan B v i gen m ha b m t HBS Ag

(Hepatitis B surface Antigen).
C Vacxin axit nucleic (vacxin ADN)

Vacxin thnh phn chnh l ADN ca plasmid ti t

hp cha gen KN.

Gen KN ca VSV gy bnh c gn vo plasmid.

a plasmid ti t hp vo trong t bo c th
bng sng bn gen,hoc vi tim.

Vacxin thng c tim vo c th a gen trc

tip vo mt s t bo c.
Khi vo trong t bo, ADN ca plasmid ti t hp
c nhn ln, protein KN c sn sinh trc tip bi
t bo vt ch, chng kch thch c th sn sinh MD.
+
u im ca vacxin ADN:
- Gen KN biu th mnh, thi gian sn xut KN lu to
MD mnh, bn, khng cn tim nhc li.
-
Vacxin ADN rt an ton v gen c c loi tr,
hiu lc n nh, d bo qun, thun tin cho vic s
dng, c ngha kinh t.
- To MD tt nhng c th b suy gim MD v c th
suy nhc.
-
C th thit k mt vacxin a gi
( Trn cng 1 plasmid gn nhiu gen m ha hoc trn
nhiu loi plasmid c cha ADN m ha cho cc loi
protein khc nhau. Hn hp vacxin ADN ny khng b
nh hng ln nhau, nh th s n gin ha c
tin trnh tim chng.
+
Nhc im

Thc nghim:
a vacxin ADN vo c th V , s phn b
ADN vacxin trong t bo khng t c mc
ti a.

Mt khc, nu ADN vacxin nhp vo h gen

ca V ch s:
- Gy hu qu v di truyn cc th h sau
- C th gy t bin t bo ung th
- c ch hot ng ca gen chng ung th,
gy bin i t bo dn n t MD.

Vacxin ADN thuc th h mi nht, c coi

l loi vacxin c trin vng ln.

Hin ti vacxin mi c nghin cu th

nghim trn V :
O
Vacxin ADN cha gen HBsAg
O
Vacxin ADN cha gen KNch yu ca HIV
O
Vacxin ADN cha gen H v N chng cm
C Vacxin xa gen c

Vacxin cha yu t gy bnh c lm nhc c

bng k thut gen ct b gen c.

V d:
O
to ra ging gc sn xut vacxin cm gia cm,
hin B Y t v Vin KHCN Vit Nam nhp ngoi
chng virus vacxin NiBRG-14 t Vin Tiu chun v
kim nghim dch sinh phm quc gia Vng quc
Anh thng qua WHO.
O
Chng NiBRG-14 to ra bng ghp 6 gen ca chng
/PR8/34 (H1N1) vi 2 gen H5N1 ca chng
A/Vietnam/1194/2004 (H5N1).
O
Ring gen H5 b loi 12 nucleotit m ha 4 a.a thuc
vng c, chng virus ny c th nui cy trn phi
g.

Vacxin Aujeski cng l 1 vacxin nhc c kiu ny.

b. Vacxin c ngun gc KN khng nhn ln
L loi vacxin v hot sn xut bng k thut gen
C Vacxin cha KN l protein sn xut bng k thut
gen

Loi vacxin ny c sn xut bng cch tch gen

KN t tc nhn gy bnh, ghp vo h gen ca mt
loi VSV lm vector dn truyn

Nui cy VSV ti t hp ny trong MT thch ng nh

h thng bioreactor hoc thit b ln men ln, lng
protein s c sn xut vi s lng nhiu sau

chit tch protein KN lm vacxin.

C Vacxin n c

Dng thc vt l bioreactor sn xut

dc cht protein l mt xu th pht trin ca
cng ngh sinh hc.
Nhiu gen m ha KN virus c chuyn vo
thc vt v biu hin vi hiu qu cao.
Sn xut vacxin n c l hng nghin
cu c trin vng
Vacxin c thc vt sn xut trong nhng
phn n c nh l, c, qu v ht.

Nm 1990 vacxin n c u tin c sn

xut t thc vt
Do 2 nh khoa hc Curtiss v
Cardineu
Cardineu khi
biu hin thnh cng protein KN b mt A ca
VK Streptococcus mutans cy thuc l.

Sau nhiu vacxin TV c cng b sn

xut trn nhiu loi cy khc nhau:
- Rau dip
- C chua
- Khoai ty, la m, u tng v ng...

Mt khc, gi l vacxin n c l cp
n s chp nhn ca c th ng vt:
. Vacxin bn vng trong dch tiu ha
. Vacxin khng b phn hy
. C kh nng kch thch p ng MD
mnh.

Ngi ta dng thut ngPlan edible

vaccine ch loi vacxin n c sn
xut t thc vt bin i gen ny.
Nguyn l sn xut vacxin n c:

Vacxin n c l loi vacxin tiu phn bao

gm mt hoc nhiu chui polypeptit ca
protein KN trong VSV gy bnh
. Ngi ta chn lc nhng gen m ha cho cc
thnh phn ny, a vo vector (plasmid hoc
vi khun ( Agrobacterium tumefaciens)
. Da vo h thng di truyn thc vt khuych
i gen v biu hin thnh cc protein KN
mong mun trong cc b phn n c ca
thc vt.
Cc thnh tu nghin cu vacxin n c
c cng b
Ngun
protein
Protein/Peptit i tng
thc vt
p ng
min dch
Tc gi, nm
bo co
E.coli
Tiu phn B c t
km chu nhit LT-
B
Thuc l
Khoai ty
KT dch th v KT
t bo theo ng
ung v n
Hagetal, 1995
Vibrio
cholerac
c t t
Tiu phn B
Khoai ty Cho n Arakawa, 1998
Virus
vim gan B
HBs-Ag Thuc l Cha th nghim Mason etal,
1992
Virus di
Glycoprotein C chua McGarvey etal
1995
Cl.tetani
Ngoi c t Lc lp
thuc l
Cho n Tregoing etal,
2003
C Vacxin peptit tng hp:

L vacxin thnh phn ch cha duy nht polypeptit

KN (8 20 axit amin)

Do ch c cc epitop KN nn khng c kh nng kch

thch sinh MD v vy sau khi cc peptit c tng hp,
ngi ta phi gn chng vo cc gi l cc ht
polyme c kh nng hp ph cao.
V d: vacxin peptit phng bnh LMLM, vic bo h t
c bng cch tim cho ng vt mt tp hp cc
chui peptit KN gm 20 axit amin.
+
Vacxin peptit c u im:
O
Sn xut v kim sot cht lng n gin.
O
Khng c cc thnh phn khng cn thit
nh nucleic, protein ngoi lai do t c.
O
C th thay i theo s bin i t nhin i
vi nhng virus khng n nh nh virus
cm.
O
C tnh kh thi thm ch trong trng hp
khng nui cy c virus.
O
n nh, gi thnh h.
+
Nhc im ca loi vacxin :
O
Sinh min dch c th km so vi cc vacxin v hot
truyn thng.
O
Bt buc phi c cht b tr.
O
Yu cu tim nhc li.
S tm tt phn loi vacxin
Vacxin
s ng
Vacxin
Vacxin v
ho t
Vacxin nh c c
ha
Vacxin s ng
nguyn c
Vacxin s ng
v c
Xa gen c
Thch ng nhi t
th p
Nh y c m nhi t

Qua v t ch trung gian

khng c m th
Ti t h p gen
Vacxin vector dn
truy n
Vacxin ADN
Vacxin d i
n v
Vacxin nguyn
ven
Vacxin
n
c
Ch a protein KN
t ng h p b ng
k thu t gen
Tinh ch
b ng k
thu t gen
Tinh ch b ng
ngu n SV ch
trong t nhin
1.6. Cht b tr
+
Khi nim:
O
Cht b tr l nhng hp cht ha hc c thm
vo trong vacxin nhm lm tng kh nng kch thch
MD v tng hiu lc ca vacxin.
O
Nm 1930, Ramon l ngi u tin nghin cu v
cht b tr. ng chng minh bng thc nghim
rng khi b sung cht b tr vo vacxin th hiu lc
ca vacxin cao hn.
* Cc cht b tr:
Gm 3 loi chnh
a. Cht b tr v c:
L nhng ht mn ca mui silicat hoc mui sunfat,
mui phosphat hoc ht bt talk, than hot tnh.
V d :
O Alumin hydroxit Al(OH)
3
O Sunfat alumin kali AlK(SO
4
)
2
.12H
2
O
O Phosphat aluminum Al(PO
4
)

Trong th y, ngi ta hay dng cht b tr l

AlK(SO
4
)
2
.12H
2
O (gi l keo phn) trong cc vacxin
vi khun v hot.
b. Cht b tr hu c:
Dng cc hp cht hu c khi trn vo vacxin s c
tnh cht huyn ph trng nh sa nn gi l nh
du.
V d: Du khong, sapolin
Du thc vt, m ng vt
Freund (du khong paraphin + BCG cht)

Nhng Freund t dng v hay gy mng m ko di

hoc vim khp.

i vi nhng KN phn t lng nh ngi ta c th

gn vi protein mang ti tng hp hoc cht mang t
nhng ht m nh gi l liposon.
.
c. B tr l sinh vt:
Thng dng l xc ca mt s loi vi khun
nh:
- M. tubercullosis
- Sal. typhimurium.
Cng c th dng ni c t ca vi khun nh
Lypopolysaccarit
* Tc dng ca cht b tr
- Kch thch MD do bn thn cht b tr gy
phn ng vim nh, ko cc i thc bo v
cc t bo MD khc ti.
- Hp ph KN, khoanh vng KN, lm chm qu
trnh gii phng KN ti v tr tim, do KN tn
ti lu trong c th, ko di s trnh din KN.
- Kch thch s hot ng ca APC qu trnh
phn tch, trnh din KN t hiu qu.
-
Cht b tr SV:
C tc dng kch thch t bo MD.
. Xc vi khun lao lm tng tng tc gia t
bo lympho v TB, do lm tng p ng
MD t bo
. LPS tc ng n t bo lympho B lm tng
qu trnh phn bo to plasmoxyte do tng
hm lng KT dch th c hiu.