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[(P3U1)-385.3(we)18.8(re)-360.4(ob)20.8(ta)19.6(in)20.7(ed)-360.5(fr)20.9(om)-360.5(th)20.7(e)-370.9(A)0(m)18.7(e)0(r)19.7(i)0(c)19.6(a)0(n)-360.5(T)0(y)19.6(p)0(e)-360.5(C)0(u)14(l)0(t)20.6(u)0(r)20.9(e)-370.9(Co)19.8(l-)]TJ
0 -1.1116 TD
[(le)19.6(ct)13.8(io)20.7(n)-289.1(\()-70.2(Ma)18.8(na)19.7(ss)15.5(as)19.9(,)-285.7(V)0(A)19.1(\))0(.)-281(H)0(S)20.1(C)0(-)19.8(2)-289.1(wa)18.8(s)-288.9(o)0(b)20.8(t)0(a)13.8(i)0(n)20.7(e)0(d)-279.8(f)0(r)20.9(o)0(m)-279.9(t)0(h)20.7(e)-290.2(Ja)19.9(pa)19.7(-)]TJ
T*
[(ne)19.7(se)-296.9(Co)14(ll)20.6(ec)18.5(ti)20.6(on)-295.9(of)-295.9(Re)18.6(se)19.9(ar)19.7(ch)-297.1(Bi)19.7(or)15.1(es)19.9(ou)20.8(rc)19.7(es)-296.9(Ce)18.6(ll)-296.2(Ba)18.6(nk)-295.9(\(O)20(sa)19.9(ka)19.7(,)]TJ
0 -1.1174 TD
[(Ja)14.1(pa)19.7(n\))20.9(.)-274.1(T)0(h)19.6(e)-278.7(LN)18.8(31)20.8(9)-271.8(c)0(e)18.5(l)0(l)-267.4(l)0(i)20.6(n)0(e)-268.3(w)0(a)18.8(s)-271.6(pr)15.1(ov)20.8(id)20.7(ed)-262.5(by)-267.1(Pr)21(of)15.1(.)-274.1(K)0(a)18.8(z)0(u)19.7(h)0(i)20.7(k)0(o)]TJ
0 -1.1116 TD
[(Mi)19.8(sh)15.3(im)19.5(a)-445.8(\()0(S)21(a)0(i)13.8(t)0(a)19.6(m)0(a)-436.6(M)0(e)18.8(d)0(i)20.7(c)0(a)18.5(l)-450.5(Un)20(iv)20.7(er)19.7(si)15.1(ty)20.7(,)-446.9(S)0(a)19.8(i)0(t)20.6(a)0(m)18.5(a)0(,)-443.4(J)0(a)19.9(p)0(a)19.7(n)0(\))20.9(.)]TJ
7.3823 0 0 6.5614 286.8661 376.044 Tm
-.0181 Tc
[(\(4)-18.1(3)6.6(\))]TJ
9.843 0 0 9.843 59.7543 360.7369 Tm
0 Tc
[(CH)18.9(O-)14.2(S)-409.9(w)0(a)18.8(s)-409.8(pu)15.1(rc)19.7(ha)19.7(se)19.9(d)-410(f)0(r)20.9(o)0(m)-406.6(T)0(h)19.6(e)0(r)19.7(m)0(o)-400.8(F)0(i)20.9(s)0(h)21(e)0(r)-406.5(S)0(c)19.8(i)0(e)19.6(n)0(t)20.7(i)0()20.9(c)0(,)-403.1(I)0(n)20.9(c)0(.)]TJ
0 -1.1174 TD
[(\(W)18.6(al)19.5(th)15(am)18.5(,)-239.6(M)0(A)19.1(\))0(.)-741.7(C)0(o)19.8(r)0(e)-233.7(f)0(u)20.9(c)0(o)19.7(s)0(e)19.9(-)0(k)15.1(n)0(o)20.8(c)0(k)19.7(o)0(u)20.8(t)-243.1(CH)18.9(O-)20(S)-242.8(c)0(e)18.5(l)0(l)20.6(s)-248.6(\(P)21(DI)20(S-)]TJ
0 -1.1116 TD
[(5\))-606.9(wa)13(s)-617.2(e)0(s)19.9(t)0(a)19.6(b)0(l)20.7(i)0(s)20.9(h)0(e)19.7(d)-617.4(us)15.3(in)20.7(g)-617.4(C)0(R)18.8(I)0(S)21(P)0(R)20(-)0(C)19.8(a)0(s)19.9(9)-617.4(in)-607.1(ou)20.8(r)-617.4(p)0(r)15.1(e)0(v)19.7(i)0(o)20.7(u)0(s)]TJ
T*
[(st)15.1(ud)20.8(y.)]TJ
7.3823 0 0 6.5614 82.9984 332.2204 Tm
-.0181 Tc
[(\(4)-18.1(1)6.6(\))]TJ
9.843 0 0 9.843 99.3826 327.855 Tm
0 Tc
[(CH)18.9(O-)20(K1)-435(an)19.7(d)-444.6(L)0(N)18.8(2)0(2)20.8(9)-444.6(ce)18.5(ll)20.6(s)-444.4(w)0(e)18.8(r)0(e)-435.3(t)0(r)20.7(a)0(n)19.7(s)0(f)15.3(e)0(c)18.5(t)0(e)19.5(d)-444.6(wi)19.8(th)]TJ
-4.026 -1.1174 TD
[(po)15.1(do)20.8(pl)20.7(an)19.7(in)-607.1(pl)20.7(as)19.9(mi)19.5(ds)-612.5(us)21(in)20.7(g)-617.4(L)0(i)19.5(p)0(o)20.8(f)0(e)19.7(c)0(t)13.8(a)0(m)24.2(i)0(n)15(e)-618.5(20)20.8(00)-607(\(T)19.7(he)19.7(rm)19.6(o)]TJ
0 -1.1116 TD
[(Fi)15.1(sh)21(er)-297.1(Sc)14.1(ie)19.5(nt)20.7(i)20.9(c,)-299.4(In)20.9(c.)17.3(,)-308.7(W)0(a)17.4(l)0(t)20.6(h)0(a)19.7(m)0(,)-299.5(M)0(A)19.1(\))-312.1(ac)18.5(co)19.7(rd)20.9(in)20.7(g)-312.1(t)0(o)-296.1(t)0(h)15(e)-307.5(ma)18.4(n-)]TJ
T*
[(uf)15.1(ac)18.5(tu)20.7(re)19.7(r)20.9(s)-271.6(i)0(n)20.7(s)0(t)20.9(r)0(u)15.1(c)0(t)19.5(i)0(o)20.7(n)0(s)21(.)]TJ
7.3823 0 0 6.5614 151.4267 299.3385 Tm
-.0181 Tc
[(\(3)-18.1(6)6.6(\))]TJ
9.843 0 0 9.843 69.7323 284.0314 Tm
0 Tc
[(The)-270.6(CHO-K1,)-267.1(CHO-K1/podoplanin,)-275(and)-269.4(P3U1)-264.3(cells)-272.8(were)]TJ
-1.0137 -1.1174 TD
[(cultured)-454.9(in)-428.5(Ro)19.8(sw)20.2(el)19.5(l)-439(P)0(a)19.8(r)0(k)-434.1(M)0(e)18.8(m)15(or)15.1(ia)19.6(l)-439(I)0(n)20.9(s)0(t)20.9(i)0(t)20.6(u)0(t)15(e)-434.2(\(R)14.1(PM)20.1(I\))-428.4(16)20.8(40)]TJ
0 -1.1116 TD
[(me)18.4(di)20.7(um)-210.8(co)13.9(nt)20.7(ai)19.5(ni)20.7(ng)]TJ
7.3823 0 0 6.5614 136.8567 260.4472 Tm
(L)Tj
9.843 0 0 9.843 141.3354 262.0912 Tm
[(-g)20.9(lu)20.7(ta)19.6(mi)19.5(ne)-210.7(\(N)20(ac)18.5(al)19.5(ai)-216.6(Te)18.5(sq)21(ue)19.7(,)-222.3(I)0(n)20.9(c)0(.)23.1(,)-222.3(Ky)20(ot)20.7(o,)]TJ
-8.2882 -1.1116 TD
[(Ja)14.1(pa)19.7(n\))-411.1(an)19.7(d)-427.3(1)0(0)20.8(%)-428.4(he)19.7(at)19.5(-i)20.7(na)19.7(ct)19.5(iv)20.7(at)13.8(ed)-412.3(fe)19.7(ta)19.6(l)-427.4(b)0(o)20.8(v)0(i)20.7(n)0(e)-418.1(s)0(e)19.9(r)0(u)20.9(m)-428.5(\(F)21(BS)20(\))]TJ
0 -1.1174 TD
[(\(T)19.7(he)13.9(rm)19.7(o)-237.2(F)0(i)15.1(s)0(h)21(e)0(r)-233.7(S)0(c)19.8(i)0(e)19.6(n)0(t)20.7(i)0()20.9(c)0(,)-236.1(I)0(n)20.9(c)0(.)23.1(\))0(.)-234.9(H)0(S)20.1(C)0(-)19.8(2)-243(ce)18.5(ll)20.6(s)-242.8(w)0(e)18.8(r)0(e)-233.7(c)0(u)19.7(l)0(t)20.6(u)0(r)20.9(e)0(d)-233.8(i)0(n)]TJ
0 -1.1116 TD
[(Ea)18.5(gl)14.9(e)19.7(s)-335(m)0(i)19.5(n)0(i)20.7(m)0(a)18.4(l)-335.3(es)14.1(se)19.9(nt)20.7(ia)19.6(l)-335.3(m)0(e)18.4(d)0(i)15(u)0(m)-326(\()0(N)20(a)0(c)18.5(a)0(l)19.5(a)0(i)-326(T)0(e)18.5(s)0(q)15.3(u)0(e)19.7(,)-337.5(In)20.9(c.)17.3(\))-335.1(o)0(r)]TJ
T*
[(Du)14.2(lb)20.7(ec)18.5(co)19.7(s)-232.4(mo)19.6(di)14.9(e)19.8(d)-243(E)0(a)18.5(g)0(l)20.7(e)0()19.7(s)-248.6(me)18.4(di)20.7(um)-233.8(\(D)20(ME)18.7(M\))-239.2(co)19.7(nt)20.7(ai)19.6(ni)20.7(ng)]TJ
7.3823 0 0 6.5614 291.1747 216.6236 Tm
(L)Tj
9.843 0 0 9.843 295.5968 218.2676 Tm
(-)Tj
-23.9604 -1.1174 TD
[(gl)14.9(ut)20.7(am)18.5(in)20.7(e)-175.1(a)0(n)19.7(d)-179.6(10)20.8(%)-175(h)0(e)19.7(a)0(t)19.5(-)0(i)15(n)0(a)19.7(c)0(t)19.5(i)0(v)20.7(a)0(t)19.5(e)0(d)-164.6(F)0(B)14.2(S)0(.)-165.6(T)0(h)19.6(e)-175(LN)18.8(31)20.8(9,)-165.8(LN)18.8(22)20.8(9,)]TJ
0 -1.1116 TD
[(an)19.7(d)-410(L)0(N)18.8(2)0(2)20.8(9)0(/)20.7(p)0(o)20.8(d)0(o)15.1(p)0(l)20.7(a)0(n)19.7(i)0(n)-394(c)0(e)18.5(l)0(l)14.8(s)-404.1(we)18.8(re)-400.7(cu)19.7(lt)20.6(ur)20.9(ed)-400.8(in)-394(DM)19.1(EM)-401.7(co)19.7(n-)]TJ
T*
[(ta)19.6(in)15(in)20.7(g)]TJ
7.3823 0 0 6.5614 92.1259 183.7417 Tm
(L)Tj
9.843 0 0 9.843 96.548 185.3858 Tm
[(-g)20.9(lu)20.7(ta)19.5(mi)19.5(ne)-544.8(an)19.7(d)-554(1)0(0)20.8(%)-555.1(he)19.7(at)19.5(-i)20.7(na)19.7(ct)19.5(iv)20.7(at)13.8(ed)-539(FB)20(S.)-545.8(Ce)18.6(ll)20.6(s)]TJ
-3.738 -1.1116 TD
[(we)18.8(re)-262.5(ke)19.7(pt)-261.5(at)-262.7(37)]TJ
/F9 1 Tf
6.1283 0 TD
()Tj
/F3 1 Tf
.3859 0 TD
[(C)-272.8(i)0(n)-261.5(a)-267.2(hu)20.8(mi)19.5(di)14.9(e)19.8(d)-271.8(a)0(t)19.5(m)0(o)19.6(s)0(p)21(h)0(e)19.7(r)0(e)-262.5(c)0(o)19.7(n)0(t)20.7(a)0(i)19.6(n)0(i)20.7(n)0(g)-261.4(5)0(%)]TJ
-6.5142 -1.1174 TD
(CO)Tj
7.3823 0 0 6.5614 73.1905 161.8015 Tm
(2)Tj
9.843 0 0 9.843 76.8189 163.4456 Tm
[(.)-245.3(A)0(l)19.8(l)-243.1(me)18.4(di)20.7(a)-244.2(c)0(o)19.7(n)0(t)20.7(a)0(i)19.5(n)0(e)19.7(d)-243(10)20.8(0)-156.6(U)0(/)14.1(m)15(L)-244.2(o)0(f)-238.3(p)0(e)19.7(n)0(i)20.7(c)0(i)19.5(l)0(l)20.6(i)0(n)20.7(,)-245.3(10)20.8(0)]TJ
/F7 1 Tf
19.871 0 TD
(m)Tj
/F3 1 Tf
.5414 0 TD
[(g/)20.7(mL)]TJ
-22.1461 -1.1116 TD
[(of)-393.8(st)20.9(re)14(pt)20.7(om)19.6(yc)19.7(in)20.7(,)-400.8(a)0(n)19.7(d)-404.3(25)]TJ
/F7 1 Tf
9.9758 0 TD
(m)Tj
/F3 1 Tf
.5414 0 TD
[(g/)20.7(mL)-396.3(of)-393.8(am)18.5(ph)20.8(ot)20.7(er)19.7(ic)19.6(in)-394(B)-405.3(\()0(N)20(a)0(c)18.5(a)0(l)19.6(a)0(i)]TJ
-10.5172 -1.1116 TD
[(Te)18.5(sq)15.3(ue)19.7(,)-268.4(I)0(n)20.9(c)0(.)17.3(\))0(.)]TJ
/F10 1 Tf
8.9663 0 0 8.9663 59.7543 116.8441 Tm
(Antibodies)Tj
/F3 1 Tf
9.843 0 0 9.843 69.7323 100.4031 Tm
[(LpMab-23,)-309.2(a)-307.5(mouse)-307.2(anti-podoplanin)-313.1(mAb)-304.9(\(IgG)]TJ
7.3823 0 0 6.5614 260.3905 98.759 Tm
(1)Tj
9.843 0 0 9.843 264.0755 100.4031 Tm
[(,)-308.7(kappa\),)]TJ
-20.758 -1.1116 TD
[(was)-310.4(developed)-313(as)-308.4(previously)-311.7(described.)]TJ
7.3823 0 0 6.5614 217.9275 93.8268 Tm
(\(42\))Tj
9.843 0 0 9.843 233.2346 89.4614 Tm
[(Human)-314(IgG)-303.6(was)]TJ
-17.6247 -1.1174 TD
[(purchased)-416.4(from)-414.5(Beckman)-416.6(Coulter,)-416.7(Inc.)-411(\(Fullerton,)-413.2(CA\))-414.1(or)]TJ
0 -1.1116 TD
[(Sigma-Aldrich)-318.4(Corp.)-319.9(\(St.)-311.9(Louis,)-315.3(MO\).)-313.8(To)-314.4(generate)-318.8(human)]TJ
T*
[(mouse)-272.7(chimeric)-278.7(anti-podoplanin)-272.8(\(chLpMab-23\),)-274.4(appropriate)]TJ
25.5097 67.9242 TD
(V)Tj
7.3823 0 0 6.5614 317.9337 723.5148 Tm
(H)Tj
9.843 0 0 9.843 326.4377 725.1588 Tm
[(and)-332.8(V)]TJ
7.3823 0 0 6.5614 351.0424 723.5148 Tm
(L)Tj
9.843 0 0 9.843 358.8094 725.1588 Tm
[(cDNAs)-331.6(of)-330.5(mouse)-330.3(LpMab-23)-334.6(and)-327(C)]TJ
7.3823 0 0 6.5614 501.3353 723.5148 Tm
(H)Tj
9.843 0 0 9.843 509.8959 725.1588 Tm
[(and)-332.8(C)]TJ
7.3823 0 0 6.5614 533.9337 723.5148 Tm
(L)Tj
9.843 0 0 9.843 541.7573 725.1588 Tm
(of)Tj
-23.4593 -1.1174 TD
[(human)-238.3(IgG)]TJ
7.3823 0 0 6.5614 355.3511 712.573 Tm
(1)Tj
9.843 0 0 9.843 361.3605 714.1605 Tm
[(were)-243.7(subcloned)-242.6(into)-240.8(pCAG-Ble)-244.3(and)-240.6(pCAG-Neo)]TJ
-5.1319 -1.1116 TD
[(vectors)-182.7(\(Wako)-181.7(Pure)-183.9(Chemical)-187.6(Industries,)-182.5(Ltd.,)-184.3(Osaka,)-184.7(Japan\),)]TJ
T*
[(respectively.)-378.6(Antibody)-378.6(expression)-375.9(vectors)-378.6(were)-376.2(transfected)]TJ
T*
[(into)-396.3(CHO-K1)-401.9(or)-399.6(PDIS-5)]TJ
7.3823 0 0 6.5614 411.5904 685.7006 Tm
(\(41\))Tj
9.843 0 0 9.843 427.8613 681.3352 Tm
[(using)-397.1(the)-402(Lipofectamine)-401.9(LTX)]TJ
-11.888 -1.1174 TD
[(reagent)-264.7(\(Thermo)-264.7(Fisher)-266.7(Scientic,)-270.3(Inc.\).)-265.8(Stable)-262.5(transfectants)]TJ
0 -1.1116 TD
[(of)-232.6(CHO-K1/chLpMab-23)-240(or)-232.6(PDIS-5/chLpMab-23)-236.9(cells)-238.3(were)]TJ
T*
[(selected)-399.4(by)-388.1(cultivating)-401(the)-390.5(transfectants)-399.1(in)-394(a)-393.9(medium)-396.3(con-)]TJ
0 -1.1174 TD
[(taining)-268.4(0.5)-166.9(mg/mL)-274.2(of)-267.1(both)-269.5(geneticin)-273(and)-269.4(zeocin)-275.2(\(InvivoGen,)]TJ
0 -1.1116 TD
[(San)-845.2(Diego,)-852(CA\).)-849.5(CHO-K1/chLpMab-23)-856.3(and)-845.4(PDIS-5/)]TJ
T*
[(chLpMab-23)-430.1(cells)-428.3(were)-428(cultivated)-429.7(in)-422.8(CHO-S-SFM)-427.8(II)-428.4(me-)]TJ
0 -1.1174 TD
[(dium)-506.9(\(Thermo)-518.1(Fisher)-508.6(Scientic,)-518(Inc.\).)-507.7(ChLpMab-23)-516.4(was)]TJ
0 -1.1116 TD
[(puried)-262.2(using)-264.6(Protein)-268.1(G-Sepharose)-263.7(\(GE)-270.3(Healthcare)-265.6(UK)-263.1(Ltd,)]TJ
T*
[(Buckinghamshire,)-284.9(England\).)]TJ
/F10 1 Tf
8.9663 0 0 8.9663 310.8472 560.7495 Tm
[(Production)-332(of)-336.7(point)-329.9(mutants)]TJ
/F3 1 Tf
9.843 0 0 9.843 320.8251 544.3085 Tm
[(The)-408.8(amplied)-415.6(human)-411.1(podoplanin)-415.6(cDNA)-411.4(was)-414.1(subcloned)]TJ
-1.0137 -1.1116 TD
[(into)-235(a)-232.6(pcDNA3)-235(vector)-239.4(\(Thermo)-235.9(Fisher)-232.1(Scientic,)-241.5(Inc.\),)-237(and)-234.9(a)]TJ
0 -1.1174 TD
[(FLAG)-220.3(epitope)-223.4(tag)-223.5(was)-224(added)-220.9(at)-222.4(the)-223.5(C-terminus.)-225.2(Amino)-220.8(acids)]TJ
0 -1.1116 TD
[(in)-382.5(podoplanin)-386.8(were)-387.7(substituted)-387.8(with)-385.7(alanine)-385.8(using)-385.6(a)-382.4(Quik-)]TJ
T*
[(Change)-585.9(Lightning)-593.2(site-directed)-589.4(mutagenesis)-592.7(kit)-591.1(\(Agilent)]TJ
0 -1.1174 TD
[(Technologies,)-349.7(Inc.,)-339.6(Santa)-347.6(Clara,)-346.5(CA\))-345(with)-345.4(oligonucleotides)]TJ
0 -1.1116 TD
[(containing)-342(the)-344.4(desired)-344(mutations.)-344.2(CHO-K1)-350.1(or)-342(CHO-S)-342.2(cells)]TJ
T*
[(were)-220.7(transfected)-224.1(with)-218.6(the)-217.7(plasmids)-218.4(using)-218.6(a)-221.1(Gene)-215(Pulser)-220.6(Xcell)]TJ
0 -1.1174 TD
[(electroporation)-378.7(system)-371.6(\(Bio-Rad)-380.7(Laboratories,)-372.7(Inc.,)-374.2(Berke-)]TJ
0 -1.1116 TD
[(ley,)-209.6(CA\).)-204.4(The)-213(point)-207.4(mutant)-205.1(cells)-209.5(of)-209.5(CHO-K1)-211.8(were)-209.2(cultured)-213(in)]TJ
T*
[(RPMI)-317.2(1640)-315.4(medium)-315.6(conta)-8(ining)]TJ
7.3823 0 0 6.5614 441.1274 433.0771 Tm
(L)Tj
9.843 0 0 9.843 445.6629 434.7211 Tm
[(-glutamine)-320.2(and)-315.5(10%)-315.4(heat-)]TJ
-13.6966 -1.1116 TD
[(inactivated)-190.1(FBS)-188.3(as)-187.5(described)-191.9(above.)-189.8(The)-190(point)-190.1(mutant)-187.8(cells)-192.2(of)]TJ
0 -1.1174 TD
[(CHO-S)-284.6(were)-278.3(cultured)-287.8(in)-278.8(CHO-S-SFMII)-285.9(medium.)]TJ
/F10 1 Tf
8.9663 0 0 8.9663 310.8472 390.8408 Tm
[(Flow)-332.3(cytometry)]TJ
/F3 1 Tf
9.843 0 0 9.843 320.8251 374.3999 Tm
[(The)-552.8(cell)-554.2(lines)-559.7(were)-554.7(harvested)-560.5(after)-555.1(brief)-559.7(exposure)-558.1(to)]TJ
-1.0137 -1.1116 TD
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T*
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T*
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T*
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T*
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(AB)Tj
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[(-23)-333.4(AGAINST)-334.7(PDPN)]TJ
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T*
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T*
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T*
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T*
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[(Cr-sodium)-444(chromate)-439.8(at)-447(37)]TJ
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(Cr-)Tj
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[(labeled)-201.5(target)-199.2(cells)-198(were)-197.6(seeded)-200(in)-198.2(96-well)-200(plates)-199.1(in)-198.2(triplicate.)]TJ
T*
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(L\))Tj
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T*
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[(is)-226.8(the)-223.5(spontaneous)]TJ
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[(Antitumor)-335.1(activity)-332.8(of)-336.7(anti-podoplanin)-338(antibodies)]TJ
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[(CHO/podoplanin)-348.1(cells)-347.7(and)-344.3(HSC-2)-347(cells)-347.7(were)-347.4(trypsinized)]TJ
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[(cells/mL)-468.8(\(CHO/podoplanin\))-471.2(or)-468.7(1.0)]TJ
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[(cells/mL)-272.9(\(HSC-2\),)-272(and)-263.7(100)]TJ
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[(L/animal)-267.4(of)-272.9(the)-263.8(cell)-271.9(suspension)]TJ
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[(L)-273(o)0(f)-267.1(1)-168.1(mg/mL)-274.2(of)-272.9(chLpMab-23)-274.6(and)-275.2(human)]TJ
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[(IgG)-332.4(were)-330.1(injected)-329.5(into)-333(the)-327.1(peritoneal)-331.6(cavity)-331.7(of)-330.5(mice.)-328.3(Addi-)]TJ
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[(cells;)-297.1(Ta-)]TJ
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T*
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[(/2,)-213.1(where)]TJ
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[(is)-215.2(the)-211.9(short)-212.8(diameter)-221.1(and)]TJ
/F8 1 Tf
9.9297 0 TD
(L)Tj
/F3 1 Tf
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[(is)-209.5(the)]TJ
-22.1864 -1.1116 TD
[(long)-407.7(diameter.)-414.6(The)-408.8(mice)-411.3(were)-416.5(euthanized)-413.3(21)-405.4(\(CHO/podo-)]TJ
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[(planin\))-285.6(or)-278.6(35)-278.7(days)-281.8(\(HSC-2\))-285.8(after)-278.6(cell)-283.5(implantation.)]TJ
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[(All)-200.3(data)-208.4(were)-203.4(expressed)-203.1(as)-204.8(means)]TJ
/F9 1 Tf
13.4259 0 TD
()Tj
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[(SEMs.)-205.4(Students)]TJ
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(-test,)Tj
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[(-test,)-384.5(one-way)-385.3(analysis)-379.6(of)-382.3(variance)-382.2(\(AN-)]TJ
-7.3897 -1.1116 TD
[(OVA\))-490.8(followed)-494.9(by)-486(TukeyKramer)-498(multiple)-489.8(comparisons,)]TJ
0 -1.1174 TD
[(and)-448(two-way)-454.2(ANOVA)-447.6(were)-451.1(performed)-451.1(as)-452.4(appropriate.)]TJ
/F8 1 Tf
23.4593 0 TD
(p-)Tj
/F3 1 Tf
-23.4593 -1.1116 TD
(Values)Tj
/F9 1 Tf
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[(0.05)-208.3(were)-214.9(considered)-216(to)-209.7(be)-210.7(signicant.)-212.6(All)-206(statistical)]TJ
-3.5422 -1.1116 TD
[(tests)-283(were)-284(two-sided.)]TJ
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8.9663 0 0 8.9663 62.022 155.2252 Tm
(Results)Tj
/F10 1 Tf
0 -1.8336 TD
[(Epitope)-332.2(mapping)-336.8(of)-336.7(LpMab-23)]TJ
/F3 1 Tf
9.843 0 0 9.843 72 122.3433 Tm
[(We)-230.3(performed)-232.2(ow)-234.4(cytometry)-232.6(using)-230.1(several)-235.7(point)-230.4(mutants)]TJ
-1.0137 -1.1174 TD
[(of)-313.2(human)-307.4(po)]TJ
5.1549 0 TD
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[(dopl)-15.9(a)9.4(n).3(in)-310.7(to)-316.5(determi)-15.9(n)10.6(e)-311.9(t).2(he)-311.9(ep)-15.9(i)10.6(t).2(ope)-311.9(o)-15.9(f)-300.3(LpMab)-15.9(-)10.7(23.)]TJ
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T*
0 Tc
[(w)15.3(e)15(re)-452.6(t)16.1(r)16.2(an)25.4(s)16.4(i)16.1(e)15(n)0(t)26.5(l)16.1(y)-461.9(e)15(xp)26.6(r)16.2(e)15(s)16.4(s)0(e)25.6(d)-461.9(i)16.1(n)-467.6(C)15.2(H)15.3(O)15.3(-)16.2(K1)-452.3(c)15(e)15(l)16.1(ls)26.7(.)-464.2(A)15.3(s)-461.7(s)0(h)26.8(o)16.2(w)15.3(n)-467.6(i)16.1(n)]TJ
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[(E57)-16(A)9.7(,)-186.5(D)-.7(58A,)-186.5(R)-.8(59)-16(A)9.7(,)-186.5(Y60A,)-186.5(o).2(r)-189.9(L)-1(64A.)-186.5(I).2(n)-189.9(c)-1(ontr)-16(a)9.5(st,)-186.5(L)-1(pM)-16(a)8.5(b-7,)]TJ
7.3823 0 0 6.5614 289.2472 71.8866 Tm
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[(\(30)6.7(\))]TJ
9.843 0 0 9.843 62.022 56.5795 Tm
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25.504 67.9242 TD
[(a)-16(l)9.3(l)-460.7(poi)-16(n)10.5(t)-460.7(mut)-16(a)9.3(nts.)-462.9(The)-16(s)9.6(e)-461.8(r).2(es)-16(u)10.8(lts)-466.2(indic)-16(a)8.3(te)-461.8(th)-16(a)9.4(t)-460.7(the)-461.8(m)-1(i)-16(n)10.5(imum)]TJ
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[(e)-15.9(p)9.5(itop)-15.9(e)-215.1(of)-224.3(Lp)-15.9(M)9.8(a)-.9(b-2)-15.9(3)-213.9(i).2(s)-224.1(G)-15.9(l)9.7(y54)-15.9(L)9.5(e)-.9(u64,)-226.7(wh)-15.9(i)10.6(c)-.9(h)-224.3(i).2(s)-229.9(v).3(ery)-230.1(s).5(imil)-15.9(a)9.4(r)-224.3(to)]TJ
0 -1.1116 TD
[(t)-15.9(h)10.6(at)-333.9(of)-328(L)-15.9(p)9.5(Mab-2)-333.8(\(Fi)-15.9(g)10.6(.)-330.4(1).3(B\))-15.9(.)-319.9(F).4(low)-334.6(c)-.9(yto)-15.9(m)9.5(etry)-333.8(and)-333.8(w)-.6(est)-15.9(e)9.4(rn)-333.8(blot)]TJ
T*
[(a)-15.9(n)9.5(alys)-15.9(e)9.7(s)-454.5(us)-15.9(i)10.8(n).3(g)-454.7(s)-15.9(e)9.7(v).3(eral)-460.6(glyc)-15.9(a)8.4(n-de)-15.9(c)9.7(ient)-460.6(podop)-15.9(l)10.6(a)-.9(nin)-460.5(cells)-460.3(re-)]TJ
T*
0 Tc
[(ve)25.4(a)15(l)16.1(e)15(d)-225.7(t)16.1(h)16.2(at)-210.8(th)26.5(e)-226.9(e)15(p)16.2(i)16.1(t)16.1(o)16.2(pe)-210.7(of)-209.5(Lp)25.4(M)15.3(a)15(b)16.2(-)16.2(2)0(3)-209.5(i)0(s)-209.5(i)0(n)26.5(d)16.2(e)15(p)16.2(e)15(n)0(d)26.6(e)15(n)16.2(t)-225.9(o)16.2(f)-225.7(g)16.2(l)16.1(yc)25.4(a)15(n)16.2(s)]TJ
0 -1.1174 TD
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[(\()-15.9(d)10.7(ata)-237(not)-236(shown)-15.9(\))10.7(,)-232.4(w)-.6(her)-15.9(e)9.6(as)-235.7(the)-231.3(e)-15.9(p)9.5(i).1(tope)-237(of)-235.8(LpMab-)-15.9(2)-219.7(i).2(nc)-15.9(l)9.4(udes)]TJ
/F8 1 Tf
23.2577 0 TD
0 Tc
(O)Tj
/F3 1 Tf
.7027 0 TD
(-)Tj
-23.9604 -1.1116 TD
-.0157 Tc
[(g)-15.7(l)10.8(ycan)-229.9(of)-229.9(Thr55)-15.7(/)10.8(S).6(er5)-15.7(6)10.9(.)]TJ
7.3823 0 0 6.5614 401.4424 663.7605 Tm
-.0162 Tc
[(\()9.6(36\))]TJ
9.843 0 0 9.843 415.3889 659.3951 Tm
-.0158 Tc
[(Here)-15.8(,)-217.3(w)-.5(e)-225.4(p).4(r)-15.8(o)10.8(duce)-15.8(d)-215(a)-231.2(human)-15.8(m)9.6(ouse)]TJ
-10.3963 -1.1116 TD
-.0159 Tc
[(c)-15.9(h)9.5(imeri)-15.9(c)-365(antib)-15.9(o)10.7(dy)-374.1(fro)-15.9(m)-359.1(L)-15.9(p)9.5(M)-.6(ab-2)-15.9(3)-363.7(a)-.9(nd)-374.1(inv)-15.9(e)9.5(stig)-15.9(a)9.5(t).2(ed)-374.1(whe)-15.9(t)9.4(her)]TJ
0 -1.1174 TD
-.0158 Tc
[(c)-15.8(h)9.6(LpMab)-15.8(-)10.8(23)-281.8(poss)-15.8(e)9.8(sse)-15.8(s)-272.4(ADCC)-282.9(a)-.8(nd)-281.8(anti)-15.8(t)10.5(u).4(mor)-281.8(act)-15.8(i)10.5(v).4(ity)-15.8(.)]TJ
/F10 1 Tf
8.9663 0 0 8.9663 313.0582 613.7006 Tm
0 Tc
[(Production)-338.3(of)-330.4(chLpMab-23)]TJ
/F3 1 Tf
9.843 0 0 9.843 323.0361 597.2596 Tm
[(We)-213(rst)-211.5(checked)-214(the)-211.9(sensitivity)-215.1(of)-209.5(chLpMab-23)-217(using)-212.8(ow)]TJ
-1.0137 -1.1116 TD
[(cytometry.)-426.1(As)-417.6(shown)-420.9(in)-417(Figure)-416.6(2A,)-421.2(chLpMab-23)-418.6(reaction)]TJ
0 -1.1174 TD
[(again)-8(st)-290.1(human)-295.9(glioblastoma)-299.3(cell)-300.7(line,)-291.5(LN319,)-299(was)-293.1(observed)]TJ
0 -1.1116 TD
[(at)-303(a)-301.8(very)-299.3(low)-303.8(concentration)-301.6(of)-301.7(10)-163.5(ng/mL.)-305.2(A)-295.7(positive)-303.9(control)]TJ
T*
[(NZ-12)-7.5(,)-493(the)-494.2(most)-495.1(sensitive)-496.1(chimeric)-503.3(anti-podoplanin)-497.4(anti-)]TJ
T*
(body,)Tj
7.3823 0 0 6.5614 335.2251 546.8597 Tm
(\(44\))Tj
9.843 0 0 9.843 351.3826 542.4944 Tm
[(reacted)-392.6(with)-385.7(LN319)-387.7(in)-388.2(the)-390.5(same)-387.9(way.)-388.9(Both)-391.5(anti-)]TJ
-3.8936 -1.1174 TD
[(bodies)-249.6(reacted)-254.4(with)-247.4(LN319)-249.5(in)-250(a)-244.2(dose-dependent)-256.1(manner.)-249.7(We)]TJ
0 -1.1116 TD
[(next)-598.9(performed)-600.9(a)-595.5(kinetic)-597.9(analy)-8(sis)-596.3(of)-595.4(the)-597.8(interaction)-597.8(of)]TJ
T*
[(chLpMab-23)-263.1(wi)19.8(th)-238.5(LN)18.8(31)20.8(9)-243(u)0(s)15.3(i)0(n)20.7(g)-243(o)21(w)-249.6(c)0(y)19.7(t)0(o)20.7(m)0(e)18.4(t)0(r)20.7(y)0(.)-240.7(A)0(s)-233.3(s)0(h)21(o)0(w)14.2(n)-243(in)]TJ
0 -1.1174 TD
[(Fi)15.1(gu)20.8(re)-170.4(2B)14(,)]TJ
/F8 1 Tf
4.3313 0 TD
(K)Tj
/F3 1 Tf
7.3823 0 0 6.5614 362.1542 496.9699 Tm
(D)Tj
9.843 0 0 9.843 369.2408 498.614 Tm
[(of)-175(ch)19.7(Lp)19.6(Ma)18.8(b-)20.9(23)-175(wa)18.8(s)-179.5(d)0(e)13.9(t)0(e)19.6(r)0(m)19.6(i)0(n)20.7(e)0(d)-170.4(t)0(o)-175.1(b)0(e)-176.2(1)0(.)24.3(2)]TJ
/F9 1 Tf
16.1157 0 TD
()Tj
/F3 1 Tf
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(10)Tj
/F9 1 Tf
7.3823 0 0 6.5614 544.5353 503.0361 Tm
(-)Tj
/F3 1 Tf
.5376 0 TD
(8)Tj
9.843 0 0 9.843 313.0582 487.6723 Tm
[(M.)-241.6(Th)19.6(e)]TJ
/F8 1 Tf
3.1678 0 TD
(K)Tj
/F3 1 Tf
7.3823 0 0 6.5614 350.7023 486.0282 Tm
(D)Tj
9.843 0 0 9.843 358.4692 487.6723 Tm
[(of)-238.3(NZ)18.8(-1)15.1(2)-248.8(w)0(a)18.8(s)-248.6(al)19.5(so)-238.2(de)19.7(te)19.6(rm)19.6(in)14.9(ed)-239.5(to)-238.5(be)-239.5(1.)18.5(2)]TJ
/F9 1 Tf
15.8392 0 TD
()Tj
/F3 1 Tf
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(10)Tj
/F9 1 Tf
7.3823 0 0 6.5614 530.9857 492.0377 Tm
(-)Tj
/F3 1 Tf
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(8)Tj
9.843 0 0 9.843 541.1337 487.6723 Tm
(M.)Tj
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0 -1.1174 TD
[(wh)14.2(ic)19.6(h)-387(p)0(o)20.8(s)0(s)21.2(e)0(s)14.1(s)0(e)19.9(s)-386.8(th)20.7(e)-393.9(h)0(i)20.7(g)0(h)20.8(e)0(s)19.9(t)-392.9(bi)20.7(nd)20.8(in)20.7(g)-387(a)0(f)19.7()0(n)15.2(i)0(t)20.6(y)]TJ
7.3823 0 0 6.5614 489.94 470.1542 Tm
[(\(8)26.6(\))]TJ
9.843 0 0 9.843 498.2739 465.7321 Tm
.3847 Tc
[(;t)384.7(h)405.4(i)384.7(ss)384.7(h)405.7(o)384.7(w)404.7(e)384.7(d)]TJ
-18.817 -1.1116 TD
0 Tc
[(th)15(at)-268.4(ch)13.9(L)15(p)0(M)14.2(a)0(b)19.7(-)0(2)20.9(3)-283.3(al)19.5(so)-272.7(po)20.8(ss)21.2(es)19.9(se)19.9(s)-283.1(h)0(i)20.7(g)0(h)-272.9(b)0(i)20.7(n)0(d)15.1(i)0(n)20.7(g)-277.6(af)14(n)21(it)20.6(y)-283.3(a)0(g)19.7(a)0(i)19.6(n)0(s)21(t)]TJ
T*
[(po)15.1(do)20.8(pl)20.7(an)19.7(in)20.7(.)-337.5(W)0(e)17.4(s)0(t)15.1(e)0(r)19.7(n)-335.2(bl)14.9(ot)-324.9(an)19.7(al)13.8(ys)21(is)-330.4(sh)21(ow)20(ed)-331.7(th)20.7(at)-331.8(ch)19.7(Lp)19.6(Ma)18.8(b-)20.9(23)]TJ
T*
[(di)14.9(d)-283.3(n)0(o)20.8(t)-289.2(de)19.7(te)19.5(ct)-274.2(T5)19.6(5A)-279.5(an)19.7(d)-289.1(S)0(5)21(6)0(A)-273.8(\()0(F)15.3(i)0(g)20.7(.)-285.7(2C)19.8(\),)-275.2(wh)20(ic)19.6(h)-289.1(i)0(s)-272.8(c)0(o)19.7(n)0(s)15.3(i)0(s)20.9(t)0(e)19.6(n)0(t)]TJ
0 -1.1174 TD
[(wi)14.1(th)-232.7(th)15(e)-244.2(r)0(e)19.7(s)0(u)15.3(l)0(t)20.6(s)-248.6(of)-232.6(o)15.2(w)-243.9(c)0(y)19.7(t)0(o)15(m)0(e)18.4(t)0(r)20.7(y)-243(us)15.3(in)20.7(g)-248.8(L)0(p)19.6(M)0(a)18.8(b)0(-)20.9(2)0(3)-238.3(\()0(F)21(i)0(g)20.7(.)-245.3(1A)20(\).)]TJ
0 -1.1116 TD
[(ch)13.9(Lp)19.6(Ma)18.8(b-)20.9(12)20.8(,)]TJ
7.3823 0 0 6.5614 365.9527 415.3322 Tm
-.0181 Tc
[(\(3)-18.1(0)6.6(\))]TJ
9.843 0 0 9.843 379.8424 410.9668 Tm
0 Tc
[(a)-192.3(h)0(u)20.8(m)0(a)18.4(n)0()20.8(m)0(o)19.6(u)0(s)15.3(e)-192.3(ch)19.7(im)19.5(er)19.7(ic)-182(an)19.7(ti)20.6(bo)15.1(dy)-180.7(pr)20.9(od)20.8(uc)19.7(ed)]TJ
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[(st)15.1(ud)20.8(y.)]TJ
7.3823 0 0 6.5614 336.3022 382.4503 Tm
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[(\(3)-18.1(7)6.6(\))]TJ
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7.3823 0 0 6.5614 522.4818 371.5085 Tm
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[(\(4)-18.1(5)6.6(\))]TJ
9.843 0 0 9.843 538.129 367.1432 Tm
0 Tc
[(an)19.7(d)]TJ
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T*
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T*
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T*
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[(chLpMab-23)-291.9(reacted)-294.7(with)-287.8(HSC-2)-295.2(in)-284.5(a)-290.2(dose-dependent)-296.5(man-)]TJ
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[(dependent)-462.8(manner.)-457.1(The)-454.9(reaction)-461.8(of)-457.2(NZ-12)-456.8(is)-457.1(much)-456.1(more)]TJ
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[(implanted)-428.6(into)-430.9(the)-430.8(anks)-426.8(of)-428.4(nude)-431.8(mice.)-432(chLpMab-23)-430.1(and)]TJ
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[(toneal)-418.1(cavity)-418.1(of)-411.1(mice,)-420.5(and)-419.2(human)-416.8(NK)-412.9(cells)-416.8(were)-422.3(injected)]TJ
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[(three)-215.3(times)-215.3(around)-215.1(the)-217.7(tumors.)-218.5(As)-216(shown)-213.5(in)-215.4(Figure)-215.1(4B,)-214(tumor)]TJ
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[(formation)-248.6(was)-247.1(observed)-252.8(in)-244.2(mice)-250(from)-247.5(the)-246.5(control)-245.2(and)-252.1(treated)]TJ
T*
[(groups.)-407.2(chLpMab-23)-418.6(signicantly)-413.1(reduced)-414.3(tumor)-410(develop-)]TJ
T*
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[(\(Fig.)-612.5(4C\).)-612.5(These)-613.6(results)-613.5(indicate)-617.5(that)-610.6(administration)-618.4(of)]TJ
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[(chLpMab-23)-303.4(with)-299.3(the)-298.3(NK)-303.4(cells)-301.6(inhibited)-299.6(the)-304.1(primary)-298.2(tumor)]TJ
T*
(growth)Tj
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[(in)-284.5(vivo)]TJ
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[(In)-520.5(a)-526.4(previous)-522.5(study,)-527.3(through)-526.2(an)-521.7(immunohistochemical)]TJ
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[(analysis,)-239.1(we)-234.6(showed)-233.1(that)-236.2(LpMab-23)-236.7(reacted)-242.8(with)-230.2(tumor)-237.2(cells)]TJ
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[(of)-486(human)-480.2(oral)-489.5(cancer)-489.3(but)-481.5(not)-487.2(with)-483.6(normal)-487.2(cells)-485.9(such)-489.2(as)]TJ
T*
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(\(42\))Tj
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[(Furthermore,)-502.8(ow)]TJ
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