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[(di)14.9(lu)20.7(te)19.6(d;)-301.8(Ag)14.2(il)20.6(en)19.7(t)-312.2(T)0(e)18.5(c)0(h)19.7(n)0(o)20.8(l)0(o)15(g)0(i)20.7(e)0(s)19.9(,)-314.4(In)20.9(c.)17.3(,)-314.4(S)0(a)19.8(n)0(t)20.7(a)-319(Cl)19.7(ar)19.7(a,)-305.2(CA)18.9(\).)-304(En)19.6(zy)19.7(-)]TJ
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[(ma)18.4(ti)14.8(c)-347.8(r)0(e)19.7(a)0(c)18.5(t)0(i)20.6(o)0(n)15.1(s)-346.5(we)18.8(re)-343.2(co)19.7(nd)20.8(uc)19.7(te)19.6(d)-352.4(u)0(s)21(i)0(n)20.7(g)-352.4(th)20.7(e)-347.8(E)0(L)18.4(I)0(S)15.3(A)-347.5(PO)20.1(D)-353.3(s)0(u)21(b)0(-)]TJ
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T*
[(me)18.4(as)14.1(ur)20.9(em)18.5(en)19.7(t)-266.2(o)0(f)-255.6(t)0(h)20.7(e)-267.2(op)20.8(ti)20.6(ca)18.5(l)-266.2(d)0(e)19.7(n)0(s)15.3(i)0(t)20.6(y)-266(at)-256.9(65)20.8(5)-156.6(n)0(m)-256.9(u)0(s)21(i)0(n)20.7(g)-266(an)-256.8(iM)19.8(ar)19.7(k)]TJ
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(C)Tj
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(9)Tj
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[(Mab-1)-7.5(1)-329.4(o)0(r)-330.5(recombinant)-339.6(C)]TJ
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(9)Tj
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[(Mab-1)-335.8(\(recC)]TJ
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(9)Tj
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(Mab-1\))Tj
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[(for)-331.5(30)]TJ
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[(minutes)-378.8(at)-377.9(4)]TJ
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()Tj
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[(of)-480.2(uorescence)-484.1(data,)-476.8(using)-483.5(SA3800)-473.9(Cell)-479.2(Analyzer)-485.6(\(Sony)]TJ
T*
[(Corp.,)-283(Tokyo,)-280.8(Japan\).)]TJ
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[(Determination)-337.6(of)-336.7(the)-334.8(binding)-332.2(afnity)-334.4(by)-332.7(ow)-335.5(cytometry)]TJ
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22.1288 0 TD
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(l)Tj
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(L)Tj
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[(4)-15.9(8)10.7(8-co)-15.9(n)10.7(juga)-15.9(t)9.4(e)-.9(d)-339.5(a)-15.9(n)9.5(t).2(i-mo)-15.9(u)10.7(s).5(e)-346.5(IgG)-346.2(\(1:20)-15.9(0)10.7(;)-339.7(C)-15.9(e)8.5(ll)-339.7(S)-15.9(i)10.7(gnal)-15.9(i)10.4(ng)-345.3(Tech-)]TJ
T*
[(n)-15.9(o)10.7(logy)-15.9(,)-250.8(Inc)-15.9(.)13(\))-264.6(was)-264.5(a)-15.9(d)9.5(d).3(ed.)-267(F).4(luor)-15.9(e)9.6(s).5(cen)-15.9(c)9.5(e)-260.1(d)-15.9(a)9.5(ta)-265.8(were)-265.8(sub)-15.9(s)10.9(eque)-15.9(n)9.5(t).1(ly)]TJ
T*
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(K)Tj
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(D)Tj
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[(\))-305.1(b).1(y)-310.9().2(ttin)-16.1(g)-294.8(t)-.1(he)]TJ
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T*
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[(Western)-336(blot)-338.1(analysis)]TJ
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[(C)-6.6(e)-1(ll)-368.6(lys)-5.4(a)-1(te)-6.8(s)-362.5(\()-5.6(10)]TJ
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6.353 0 TD
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(l)Tj
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[(g\))-362.6(w)-6.4(e)-.9(re)-369.5(boi)-5.6(led)-368.4(i).1(n)-362.6(s)-5.3(od)-5.5(ium)-369.6(dod)-5.5(ecy)-5.5(l)-362.7(s).4(u)-5.5(l).1(f)-5.5(a)-.9(te)]TJ
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0 -1.1116 TD
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T*
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0 -1.1174 TD
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T*
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12.5158 0 TD
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[(embranes)-257.2(w)-4.8(e).7(re)-264.3(incubate)-5.1(d)-257.3(w)1(ith)]TJ
-12.5158 -1.1116 TD
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(l)Tj
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[(Mab-11)-356(and)-350.2(1)]TJ
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[(g/mL)-351.1(of)-355.6(an)-349.9(anti-DY)-5.2(KDD)-5.2(DDK)]TJ
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T*
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[(g/)-5.4(mL)-236.9(of)]TJ
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(24,25)Tj
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()Tj
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/F8 1 Tf
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[(TANAKA)-335.9(ET)-338.7(AL.)]TJ
ET
endstream
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BT
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(Results)Tj
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[(Establishment)-337.8(of)-336.7(anti-hCCR9)-332.2(mAbs)]TJ
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[(We)-213(rst)-211.5(immunized)-210.8(mice)-209.7(with)-212.9(the)-211.9(hCCR9)-210.2(peptide)-211.9(\(Fig.)-209.4(1\).)]TJ
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[(Hybridomas)-457.7(were)-451.1(seeded)-453.4(into)-453.9(96-well)-453.5(plates,)-450.2(after)-457.1(which)]TJ
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[(ELISA)-262(was)-258.6(used)-258.8(to)-261.5(select)-263.6(reactive)-261.4(wells)-261(for)-256.7(hCCR9)-262(peptide,)]TJ
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[(followed)-517.9(by)-514.8(the)-511.4(selection)-520.4(of)-509(CHO/h)-8.4(CCR9)-514.4(and)-511.3(MOLT-4)]TJ
T*
[(cells-reactive)-333.7(supern)-7.5(atants)-325.8(of)-324.7(hybridoma)-329.2(using)-328(ow)-326.6(cytom-)]TJ
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25.5097 8.8469 TD
[(\(6.58%\),)-440.7(and)-442.2(nally)-445.6(established)-447.6(C)]TJ
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(9)Tj
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[(Mab-11)-440.6(\(mouse)-446.5(IgG)]TJ
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(2a)Tj
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(,)Tj
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[(Flow)-332.3(cytometry)]TJ
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(9)Tj
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[(Mab-11)-601.9(and)]TJ
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(recC)Tj
7.3823 0 0 6.5614 329.4991 658.0344 Tm
(9)Tj
9.843 0 0 9.843 333.1842 659.6786 Tm
[(Mab-1)-796.6(against)-799.2(exogenously)-797.7(hCCR9-overexpressed)]TJ
-2.2693 -1.1174 TD
[(CHO/hCCR9)-240.6(and)-234.9(endogenously)-240.1(hCCR9-expressed)-247.3(MOLT-4)]TJ
0 -1.1116 TD
[(cells.)-1215.1(Results)-1219.3(showed)-1212.2(that)-1215.4(C)]TJ
7.3823 0 0 6.5614 459.7227 636.0943 Tm
(9)Tj
9.843 0 0 9.843 463.4644 637.7384 Tm
[(Mab-11)-1212.4(recognized)]TJ
ET
q
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W n
q
324.113 0 0 519.534 142.809 93.316 cm
/Im2 Do
Q
Q
BT
/F7 1 Tf
9.845 0 0 9.845 59.7543 76.5354 Tm
[(FIG.)-302.2(1.)]TJ
/F4 1 Tf
4.0828 0 TD
[(A)-295.5(schematic)-302.9(illustration)-301.9(about)-294.2(the)-303.8(production)-299.3(of)-295.7(anti-hCCR9)-302.3(mAbs.)-297.1(The)-299(mice)-301.3(were)-300.8(intraperitoneally)-299(immunized)]TJ
-4.0828 -1.0135 TD
[(with)-235.5(the)-240.4(hCCR9)-232.5(peptide.)-237.6(Screening)-237(of)-238.1(hybridoma)-236(was)-235.1(then)-235.6(conducted)-237.2(by)-232.3(ELISA,)-241.6(followed)-234.8(by)-232.3(ow)-239.7(cytometry)-237.3(using)-229.5(hCCR9-)]TJ
T*
[(expressing)-340.3(cells.)-333.3(ELISA,)-333.8(enzyme-linked)-341.5(immunosorbent)-333.3(assay;)-336.3(mAbs,)-337.4(monoclonal)-336.2(antibodies;)-336.2(hCCR9,)-333.8(human)-335.5(CCR9.)]TJ
/F8 1 Tf
8.9663 0 0 8.9663 59.7543 749.7069 Tm
[(DEVELOPMENT)-339.9(OF)-337.5(ANTI-)]TJ
6.4556 0 0 6.4556 170.4189 749.7069 Tm
(H)Tj
8.9663 0 0 8.9663 175.0677 749.7069 Tm
(CCR9)Tj
6.4556 0 0 6.4556 202.3369 749.7069 Tm
(M)Tj
8.9663 0 0 8.9663 207.7228 749.7069 Tm
(AB)Tj
36.5085 0 TD
(305)Tj
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