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[(Fluorescenc)-10.8(e)-555.2(data)-554(were)-560.5(collected)-556.4(using)-558.4(the)-557.5(EC800)-554.9(Cell)]TJ
T*
[(Analyzer)-289.7(\(Sony)-276(Corp.,)-283(Tokyo,)-280.8(Japan\).)]TJ
/F9 1 Tf
8.9663 0 0 8.9663 313.0582 533.3101 Tm
[(Determination)-337.6(of)-336.7(binding)-332.2(afnity)]TJ
/F4 1 Tf
9.843 0 0 9.843 323.0361 516.8691 Tm
[(SNP)-412.8(cells)-416.8(were)-416.5(suspended)-415(in)-411.3(100)]TJ
/F10 1 Tf
14.2956 0 TD
(l)Tj
/F4 1 Tf
.5472 0 TD
[(L)-417(o)0(f)-411.1(serially)-414.5(diluted)]TJ
-15.8565 -1.1116 TD
[(E134Bf)-475.3(\(0.00610)-6.5(0)]TJ
/F10 1 Tf
8.3401 0 TD
(l)Tj
/F4 1 Tf
.5529 0 TD
[(g/mL\))-474.6(followed)-477.6(by)-474.5(FITC-conjugated)]TJ
-8.893 -1.1116 TD
[(anti-dog)-217.6(IgG)-211.5(\(1:200\).)-212.8(Fluorescence)-214.6(data)-214.2(were)-214.9(collected)-216.5(using)]TJ
0 -1.1174 TD
[(the)-200.4(EC800)-192.1(Cell)-196.9(Analyzer.)-201(The)-195.7(dissociation)-194.2(constant)-201.3(\()]TJ
/F11 1 Tf
21.0056 0 TD
(K)Tj
/F4 1 Tf
7.3823 0 0 6.5614 526.3936 482.3432 Tm
(D)Tj
9.843 0 0 9.843 531.7227 483.9873 Tm
[(\))-191.1(was)]TJ
-22.2152 -1.1116 TD
[(calculated)-426.2(by)-422.7(tting)-421.5(binding)-416.9(isotherms)-421.1(to)-422.8(built-in)-418.2(one-site)]TJ
T*
[(binding)-365.1(models)-360.3(in)-359.4(GraphPad)-363.2(Prism)-363.6(8)-358.2(\(GraphPad)-364.2(Software,)]TJ
0 -1.1174 TD
[(Inc.,)-282(La)-286.8(Jolla,)-280.9(CA\).)]TJ
/F9 1 Tf
8.9663 0 0 8.9663 313.0582 423.7227 Tm
(ADCC)Tj
/F4 1 Tf
9.843 0 0 9.843 323.0361 407.2818 Tm
[(Canine)-499.7(mononuclear)-500.8(cells)-497.5(\()-87.5(MNC\))-496.8(were)-497.1(obtained)-501(from)]TJ
-1.0137 -1.1116 TD
[(Yama)-8.8(guchi)-738.3(University)-744.7(and)-735.9(resuspended)-746.7(in)-739.6(Dulbeccos)]TJ
0 -1.1174 TD
[(modied)-194.4(Eagles)-195.4(medium)-188.9(\(DMEM\))-194.7(with)-189.8(10%)-188.7(FBS)-194.1(to)-186.6(be)-193.4(used)]TJ
0 -1.1116 TD
[(as)-665.5(effector)-668.9(cells.)-667.9(Target)-666.8(SNP)-666.2(cells)-664.5(were)-669.9(labeled)-668.1(with)]TJ
T*
(10)Tj
/F10 1 Tf
1.1692 0 TD
(l)Tj
/F4 1 Tf
.5529 0 TD
[(g/mL)-335.3(Calcein)-340.8(AM)-338(\(Thermo)-339.5(Fisher)-335.8(Scientic,)-339.4(Inc.\))-343(and)]TJ
-1.7222 -1.1174 TD
[(resuspended)-372.3(in)-370.9(the)-367.5(same)-370.6(medium.)-365.1(The)-368.5(target)-372(cells)-370.8(\(2)]TJ
/F8 1 Tf
22.1979 0 TD
()Tj
/F4 1 Tf
.72 0 TD
(10)Tj
7.3823 0 0 6.5614 548.5038 356.8818 Tm
(4)Tj
9.843 0 0 9.843 313.0582 341.518 Tm
[(cells/well\))-304.9(were)-295.6(plated)-297.2(in)-296.1(96-well)-298(plates)-297(and)-298.2(mixed)-296(with)-293.5(ef-)]TJ
0 -1.1116 TD
[(fector)-481.3(cells)-474.4(\(effector/target)-484.6(cells)-474.4(ratio,)-476.9(50\),)-479(100)]TJ
/F10 1 Tf
20.2569 0 TD
(l)Tj
/F4 1 Tf
.5529 0 TD
[(g/mL)-473.5(of)]TJ
-20.8098 -1.1174 TD
[(E134Bf,)-185(or)-180.7(control)-181.9(dog)-181.9(IgG.)-180.4(Following)-183.8(4.5-hour)-183.9(incubation)-180.8(at)]TJ
0 -1.1116 TD
(37)Tj
/F8 1 Tf
1.0022 0 TD
()Tj
/F4 1 Tf
.3974 0 TD
[(C,)-339.6(the)-344.4(release)-346.3(of)-342(Calcein)-346.6(into)-338.7(the)-344.4(supernatant)-344(was)-345(mea-)]TJ
-1.3996 -1.1116 TD
[(sured)-455.7(in)-445.8(each)-451.4(well.)-453.6(The)-449.2(uorescence)-455.3(intensity)-454.9(was)-448.6(deter-)]TJ
T*
[(mined)-365.1(using)-362.5(a)-365.1(microplate)-367.4(reader)-361.4(\(Power)-365.5(Scan)-363.7(HT;)-362.6(BioTek)]TJ
0 -1.1174 TD
[(Instruments,)-374.9(Inc.,)-368.4(Winooski,)-371.7(VT\))-368.2(with)-368.4(an)-366.2(excitation)-372.1(wave-)]TJ
0 -1.1116 TD
[(length)-284.5(of)-278.6(485)-170.3(nm)-279.9(and)-280.9(an)-279.8(emission)-281.8(wavelength)-286.3(of)-278.6(538)-170.3(nm.)]TJ
1.0137 -1.1116 TD
[(Cytolytic)-329.4(activity)-328.4(\(%)-325.8(lysis\))-329(was)-327.7(calculated)-328.2(as)-325.7(follows:)-326.6(%)]TJ
-1.0137 -1.1174 TD
(lysis)Tj
/F8 1 Tf
2.0044 0 TD
(=)Tj
/F4 1 Tf
.72 0 TD
(\(E)Tj
/F8 1 Tf
1.2038 0 TD
(-)Tj
/F4 1 Tf
.8121 0 TD
[(S\)/\()-84.9(M)]TJ
/F8 1 Tf
2.7243 0 TD
(-)Tj
/F4 1 Tf
.8121 0 TD
(S\))Tj
/F8 1 Tf
1.054 0 TD
()Tj
/F4 1 Tf
.72 0 TD
[(100,)-254.4(where)-262.1()45(E)53.2()-260.2(i)0(s)-261.3(the)-258(uorescence)]TJ
-10.0507 -1.1116 TD
[(measured)-347.4(in)-342.1(combined)-346.6(cultures)-345.2(of)-342(target)-349(and)-338.5(effector)-346.4(cells,)]TJ
T*
[()45(S)54.5()-248.7(i)0(s)-244(the)-246.5(spontaneous)-250.3(uorescence)-247.9(of)-244.1(target)-251.1(cells)-244(only,)-249.9(and)]TJ
0 -1.1174 TD
[()45(M)53.5()-317.8(i)0(s)-318.9(the)-321.4(maximum)-318(uorescence)-322.8(measured)-324.3(following)-322.2(the)]TJ
0 -1.1116 TD
[(lysis)-247.3(of)-249.8(all)-246.6(cells)-249.8(with)-247.4(a)-244.2(buffer)-249.6(containing)-244.1(0.5%)-249.7(Triton)-244.2(X-100,)]TJ
T*
[(10)-169.2(mM)-280.8(Tris-HCl)-282.6(\(pH)-280.6(7.4\),)-280.8(and)-280.9(10)-169.2(mM)-280.8(of)-278.6(EDTA.)]TJ
/F9 1 Tf
8.9663 0 0 8.9663 313.0582 160.6676 Tm
(CDC)Tj
/F4 1 Tf
9.843 0 0 9.843 323.0361 144.2267 Tm
[(SNP)-372.5(cells)-370.8(were)-370.4(labeled)-374.3(with)-374.2(10)]TJ
/F10 1 Tf
13.3164 0 TD
(l)Tj
/F4 1 Tf
.5529 0 TD
[(g/mL)-369.8(Calcein)-375.4(AM)-372.6(and)]TJ
-14.8831 -1.1116 TD
[(resuspended)-199.5(in)-198.2(the)-194.7(medium.)-198.1(They)-191.1(were)-197.6(then)-195.8(plated)-199.3(in)-192.4(96-well)]TJ
T*
[(plates)-291.2(at)-285.7(2)]TJ
/F8 1 Tf
4.314 0 TD
()Tj
/F4 1 Tf
.7142 0 TD
(10)Tj
7.3823 0 0 6.5614 372.4156 126.7086 Tm
(4)Tj
9.843 0 0 9.843 378.9353 122.3433 Tm
[(cells/well)-286.5(with)-287.8(rabbit)-290.2(complement)-287.9(\(nal)-283.1(di-)]TJ
-6.6928 -1.1174 TD
[(lution)-536.9(1:10;)-535.7(Low-Tox-M)-532.4(Rabbit)-539(Complement;)-536.7(Cedarlane)]TJ
0 -1.1116 TD
[(Labor)-7.8(atories,)-243.9(Hornby,)-241.3(Canada\))-248.4(and)-240.6(100)]TJ
/F10 1 Tf
16.0581 0 TD
(l)Tj
/F4 1 Tf
.5529 0 TD
[(g/mL)-243.1(of)-238.3(E134Bf)-245(or)]TJ
-16.611 -1.1116 TD
[(control)-210.7(dog)-199.1(IgG.)-203.4(Following)-206.8(4)-202.7(hours)-206.9(of)-198(incubation)-209.6(at)-205.1(37)]TJ
/F8 1 Tf
21.6104 0 TD
()Tj
/F4 1 Tf
.3974 0 TD
[(C,)-201.4(we)]TJ
-22.0078 -1.1174 TD
[(measured)-278.3(Calcein)-277.4(release)-277.2(into)-269.6(the)-275.3(supernatant)-274.8(for)-273.9(each)-272.8(well.)]TJ
0 -1.1116 TD
[(Fluorescenc)-10.8(e)-457.3(intensity)-460.7(was)-465.9(calculated)-460.7(as)-463.9(described)-462.6(in)-463.1(the)]TJ
T*
[(ADCC)-284.7(section)-286.6(above.)]TJ
/F6 1 Tf
8.9663 0 0 8.9663 62.022 749.6502 Tm
(54)Tj
50.2102 0 TD
[(LI)-331.6(ET)-338.7(AL.)]TJ
ET
endstream
endobj
35 0 obj
<>stream
/GS1 gs
BT
/F9 1 Tf
8.9663 0 0 8.9663 59.7543 725.7825 Tm
0 0 0 rg
0 Tc
0 Tw
[(Antitumor)-290.8(activity)-282.2(of)-286.2(E134Bf)-284.5(in)-285.9(xenografts)-283.2(of)-286.2(SNP)-289.5(cells)]TJ
/F4 1 Tf
9.843 0 0 9.843 69.7323 709.3416 Tm
[(A)-405.1(total)-404.5(of)-399.6(16)-405.4(female)-408.8(BALB/c)-403.7(nude)-403(mice)-405.5(\(5)-405.3(weeks)-405.9(old,)]TJ
-1.0137 -1.1116 TD
[(weighing)-350.9(1417)-166.8(g\))-342(were)-353.2(purchased)-347.2(from)-345.4(Charles)-350.8(River)-341.9(La-)]TJ
0 -1.1174 TD
[(boratories,)-270.2(Inc.)-267.1(\(Kanagawa,)-267.4(Japan\),)-271.3(and)-263.6(used)-264.6(in)-267.3(experiments)]TJ
0 -1.1116 TD
[(once)-444.5(they)-443.4(reached)-444.3(7)-444.6(weeks)-446.2(of)-439.9(age.)-441(SNP)-441.6(cells)-445.6(\(0.3)-168(mL)-442.4(of)]TJ
T*
(1.33)Tj
/F8 1 Tf
1.918 0 TD
()Tj
/F4 1 Tf
.72 0 TD
(10)Tj
7.3823 0 0 6.5614 95.5842 669.8833 Tm
(8)Tj
9.843 0 0 9.843 101.8204 665.5179 Tm
[(cells/mL)-267.2(in)-267.3(DMEM\))-262.8(were)-266.8(mixed)-267.2(with)-259(0.5)-166.9(mL)-263.8(BD)]TJ
-4.2737 -1.1174 TD
[(Matrigel)-375.3(Matrix)-371.8(Growth)-371.3(Factor)-371.7(Reduced)-375.1(\(BD)-368(Biosciences,)]TJ
0 -1.1116 TD
[(San)-280.8(Jose,)-279.3(CA\);)-277.1(100)]TJ
/F10 1 Tf
7.9714 0 TD
(l)Tj
/F4 1 Tf
.5529 0 TD
[(L)-278.8(o)0(f)-272.9(this)-280.9(suspension)-282.4(\(5)]TJ
/F8 1 Tf
9.3883 0 TD
()Tj
/F4 1 Tf
.72 0 TD
(10)Tj
7.3823 0 0 6.5614 253.0204 647.9431 Tm
(6)Tj
9.843 0 0 9.843 259.4267 643.5778 Tm
[(cells\))-279.7(was)]TJ
-20.2857 -1.1116 TD
[(injected)-289.2(subcutaneously)-283.7(into)-281.1(the)-281.1(left)-282.3(anks)-282.8(of)-278.6(the)-281.1(mice.)]TJ
1.0137 -1.1116 TD
[(On)-452.3(day)-453.7(6)-456.1(postinoculation,)-454.7(100)]TJ
/F10 1 Tf
13.0861 0 TD
(l)Tj
/F4 1 Tf
.5472 0 TD
[(g)-456.1(o)0(f)-451.4(E134Bf)-452.3(\()]TJ
/F11 1 Tf
6.1398 0 TD
(n)Tj
/F8 1 Tf
.6681 0 TD
(=)Tj
/F4 1 Tf
.72 0 TD
[(8\))-451.4(or)]TJ
-22.1749 -1.1174 TD
[(co)13.9(nt)20.7(ro)20.9(l)-387.1(d)0(o)15.1(g)-387(Ig)20.9(G)-393.6(\()]TJ
/F11 1 Tf
7.2918 0 TD
(n)Tj
/F8 1 Tf
.6566 0 TD
(=)Tj
/F4 1 Tf
.7084 0 TD
[(8\))-376.5(in)-382.5(10)20.8(0)]TJ
/F12 1 Tf
4.0088 0 TD
(m)Tj
/F4 1 Tf
.5414 0 TD
[(L)-388.2(P)0(B)14.2(S)-386.8(we)18.8(re)-383.5(in)20.7(je)19.6(ct)19.6(ed)-383.5(in)20.7(tr)20.7(a-)]TJ
-13.207 -1.1116 TD
[(pe)13.9(ri)20.7(to)20.7(ne)19.7(al)19.5(ly)20.7(.)-182(A)0(d)20(d)0(i)15(t)0(i)20.6(o)0(n)20.8(a)0(l)-170.5(a)0(n)19.7(t)0(i)20.6(b)0(o)15.1(d)0(y)-169.2(i)0(n)20.7(o)0(c)19.7(u)0(l)20.7(a)0(t)13.8(i)0(o)20.7(n)0(s)-169(w)0(e)18.8(r)0(e)-170.4(p)0(e)13.9(r)0(f)20.9(o)0(r)20.9(m)0(e)18.4(d)]TJ
T*
[(on)-226.8(da)19.7(ys)-226.6(15)-221.1(an)19.7(d)-231.5(2)0(3)15.1(.)-233.8(Ca)18.6(ni)20.7(ne)-222.2(MN)19.1(C)-238.3(\()0(6)20.9(.)0(3)]TJ
/F8 1 Tf
14.837 0 TD
()Tj
/F4 1 Tf
.7084 0 TD
(10)Tj
7.3823 0 0 6.5614 222.4062 593.1778 Tm
(5)Tj
9.843 0 0 9.843 228.4157 588.8124 Tm
[(ce)18.5(ll)20.6(s\))21.1(,)-233.8(w)0(h)20(i)0(c)13.8(h)-231.5(we)18.8(re)]TJ
-17.1351 -1.1174 TD
[(ob)15.1(ta)19.5(in)20.7(ed)-573.6(fr)20.9(om)-573.6(Ya)18.8(ma)18.4(gu)15.1(ch)19.7(i)-583(U)0(n)20(i)0(v)20.7(e)0(r)19.7(s)0(i)15.1(t)0(y)20.7(,)-585.2(we)18.8(re)-573.5(in)20.7(je)19.6(ct)19.5(ed)-579.3(su)21(r-)]TJ
0 -1.1116 TD
[(ro)15.1(un)20.8(di)20.7(ng)-203.8(th)20.7(e)-215.4(t)0(u)20.7(m)0(o)19.6(r)0(s)-203.6(t)0(r)20.7(e)0(a)18.5(t)0(e)19.6(d)-214.2(wi)19.8(th)-203.9(bo)20.8(th)-203.9(do)20.8(g)-214.2(I)0(g)20.9(G)-215.1(an)19.7(d)-214.2(E)0(1)19.6(3)0(4)20.8(B)0(f)-204.8(o)0(n)]TJ
T*
[(da)13.9(ys)-226.6(6,)-229.2(15)20.8(,)-239.6(a)0(n)19.7(d)-237.2(23)15.1(.)-233.8(A)0(t)-233.6(2)0(8)-226.8(d)0(a)19.7(y)0(s)-232.4(f)0(o)20.9(l)0(l)20.6(o)0(w)20(i)0(n)20.7(g)-243(ce)18.5(ll)-227.1(im)19.5(pl)20.7(an)13.9(ta)19.6(ti)20.6(on)20.8(,)-239.6(a)0(l)19.6(l)]TJ
0 -1.1174 TD
[(mi)19.5(ce)-292.5(we)18.8(re)-291.3(eu)19.7(th)20.7(an)13.9(iz)19.5(ed)-285.6(by)-290.2(ce)18.5(rv)15.1(ic)19.6(al)-291.5(di)20.7(sl)20.9(oc)19.7(at)19.5(io)15(n.)-286.8(Tu)19.6(mo)19.6(r)-300.6(d)0(i)20.7(a)0(m)18.5(e)0(-)]TJ
0 -1.1116 TD
[(te)13.8(rs)-215.1(an)19.7(d)-225.7(v)0(o)20.8(l)0(u)20.7(m)0(e)18.4(s)-225.5(we)18.8(re)-216.4(de)13.9(te)19.6(rm)19.6(in)20.7(ed)-216.5(as)-216.3(pr)20.9(ev)13.9(io)20.7(us)21(ly)-215.4(de)19.7(sc)19.9(ri)15(be)19.7(d.)]TJ
7.3823 0 0 6.5614 286.8661 538.3558 Tm
-.0181 Tc
[(\(1)-18.1(3)6.6(\))]TJ
/F9 1 Tf
8.9663 0 0 8.9663 59.7543 511.8802 Tm
0 Tc
[(Statistical)-336.1(analyses)]TJ
/F4 1 Tf
9.843 0 0 9.843 69.7323 495.4392 Tm
[(All)-482.5(data)-479.2(are)-483.6(expressed)-485.3(as)-481.2(mean)]TJ
/F8 1 Tf
13.7139 0 TD
()Tj
/F4 1 Tf
.72 0 TD
[(standard)-483.3(error)-478.9(of)-480.2(the)]TJ
-15.4475 -1.1116 TD
[(mean.)-368.5(Statistical)-361.6(analysis)-368.1(was)-362.2(conducted)-364.9(with)-362.6(Tukeys)-361.2(test)]TJ
T*
[(for)-360.3(ADCC)-365.3(and)-361.6(CDC,)-361(and)-355.8(Welchs)]TJ
/F11 1 Tf
14.6354 0 TD
(t)Tj
/F4 1 Tf
.6451 0 TD
[(test)-357(for)-360.3(tumor)-363.9(weight.)]TJ
-15.2805 -1.1174 TD
[(Analysis)-275.7(of)-272.9(variance)-278.5(and)-275.2(Sidaks)-273.6(multiple)-270.9(comparisons)-280.3(tests)]TJ
0 -1.1116 TD
[(were)-422.3(conducted)-416.7(for)-417.9(tumor)-415.8(volume)-418.1(and)-413.4(mouse)-416.6(weight.)-415.5(All)]TJ
T*
[(calculations)-732.4(were)-733.3(performed)-727.6(using)-731.2(GraphPad)-731.8(Prism)-726.4(8)]TJ
T*
[(\(GraphPad)-335.4(Software,)-326.4(Inc.\).)-329.1(A)]TJ
/F11 1 Tf
12.3431 0 TD
(p)Tj
/F4 1 Tf
.4953 0 TD
[(-value)-331.6(of)]TJ
/F8 1 Tf
3.9972 0 TD
(<)Tj
/F4 1 Tf
.5529 0 TD
[(0.05)-329.3(was)-327.7(consid-)]TJ
-17.3886 -1.1174 TD
[(ered)-283.2(statistically)-289(signicant.)]TJ
/F6 1 Tf
8.9663 0 0 8.9663 59.7543 396.6802 Tm
(Results)Tj
/F9 1 Tf
0 -1.8336 TD
[(Flow)-338.6(cytometry)-334.3(analysis)-334.5(against)-333.8(a)-333.2(canine)-330.3(mammary)]TJ
0 -1.2266 TD
[(gland)-337.1(tumor)-332.3(cell)-333(line,)-335.1(SNP)-340.1(cells)-332.5(using)-332.2(E134Bf)]TJ
/F4 1 Tf
9.843 0 0 9.843 69.7323 352.7999 Tm
[(In)-376.5(our)-371.9(previous)-378.5(study,)-377.5(an)-372(anti-hEGFR)-378.2(mAb,)-377.5(EMab-134,)]TJ
-1.0137 -1.1116 TD
[(recognized)-344.1(dEGFR-overexpressed)-344.8(CHO/dEGFR)-338.3(cells,)-339.6(indi-)]TJ
T*
[(cating)-435.4(that)-437.8(EMab-134)-432.5(crossreacts)-436.7(with)-437.5(dEGFR.)]TJ
7.3823 0 0 6.5614 255.5716 335.2818 Tm
(\(13\))Tj
9.843 0 0 9.843 272.1826 330.9164 Tm
[(In)-428.4(this)]TJ
-21.5816 -1.1174 TD
[(study,)-371.7(we)-367.1(produced)-374(the)-367.5(defucosylated)-370.4(mouse-dog)-369.2(chimeric)]TJ
0 -1.1116 TD
[(mAb)-644.7(from)-639.1(EMab-134)-639.9(\(E134Bf)-84.8(\))-640.4(and)-638(demonstrated)-645.8(that)]TJ
T*
[(E134Bf)-227.7(detected)-226.8(SNP)-228.5(cells)-226.8(\(Fig.)-220.9(1A\),)-226.4(indicating)-227(that)-224.7(E134Bf)]TJ
0 -1.1174 TD
[(detect)-286.8(endogenous)-283.9(dEGFR)-279.2(expressed)-283.7(on)-284.4(SNP)-280.3(cells.)]TJ
/F9 1 Tf
8.9663 0 0 8.9663 59.7543 264.9826 Tm
[(Determination)-337.6(of)-336.7(the)-328.5(binding)-338.5(afnity)]TJ
/F4 1 Tf
9.843 0 0 9.843 69.7323 248.5417 Tm
[(A)-255.4(kinetic)-258.1(analysis)-258.7(of)-255.6(the)-252.3(interactions)-258.9(of)-255.6(E134Bf)-256.5(with)-253.2(SNP)]TJ
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[(cells)-618.4(was)-615.7(performed)-618.1(via)-609.4(ow)-614.5(cytometry.)-616.1(As)-619.2(shown)-611(in)]TJ
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[(Figure)-359(1B,)-358(the)]TJ
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(K)Tj
/F4 1 Tf
7.3823 0 0 6.5614 128.6929 224.9574 Tm
(D)Tj
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[(for)-360.3(the)-355.9(interaction)-361.7(of)-359.3(E134Bf)-360.2(with)-356.9(SNP)]TJ
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[(cells)-290.1(was)-287.4(9.7)]TJ
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()Tj
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(10)Tj
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(-)Tj
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(10)Tj
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[(M,)-287.7(suggesting)-288.5(that)-288.1(E134Bf)-285.3(shows)-288.2(high)]TJ
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[(afnity)-285.4(for)-279.7(SNP)-280.3(cells.)]TJ
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[(E134Bf-mediated)-336.1(ADCC)-335.1(and)-335.7(CDC)-337.2(in)-330.2(SNP)-333.7(cells)]TJ
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9.843 0 0 9.843 69.7323 166.1669 Tm
[(We)-287.9(next)-287.9(investigated)-293.5(whether)-287.5(E134Bf)-291(has)-286.5(ADCC)-290.4(against)]TJ
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[(SNP)-545.3(cells)-543.5(using)-541.1(canine)-540.1(MNC)-541.8(as)-544.6(effector.)-545.7(As)-538.5(shown)-541.8(in)]TJ
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[(Figure)-537.6(2A,)-542.2(E134Bf)-532.9(showed)-538.3(ADCC)-543.9(\(37.0%)-534.1(cytotoxicity\))]TJ
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[(against)-390.2(SNP)-384(cells)-382.3(more)-386.9(potent)-382.4(than)-385.8(dog)-383.4(IgG)-384.3(\(14.4%)-384.4(cyto-)]TJ
T*
[(toxicity\))-282.3(and)-286.7(PBS)-280.5(\(10.2%)-280.7(cytotoxicity\).)]TJ
1.0137 -1.1174 TD
[(We)-391.5(then)-391.6(investigated)-391.4(whether)-396.9(E134Bf)-389(has)-390.2(CDC)-392.1(against)]TJ
-1.0137 -1.1116 TD
[(SNP)-280.3(cells.)-276.3(As)-279.3(shown)-276.9(in)-273(Figure)-278.4(2B,)-277.4(E134Bf)-279.5(elicited)-277.8(a)-278.7(higher)]TJ
T*
[(degree)-286.5(of)-284.4(CDC)-288.5(\(37.7%)-286.4(cytotoxicity\))-288(in)-278.8(SNP)-286.1(cells)-290.1(compared)]TJ
0 -1.1174 TD
[(with)-305(that)-299.6(elicited)-306.6(by)-301.7(dog)-302.8(IgG)-297.9(\(14.4%)-303.7(cytotoxicity\))-305.3(and)-298.2(PBS)]TJ
0 -1.1116 TD
[(\(9.4%)-245(cytotoxicity\).)-245.3(These)-245(results)-239.1(demonstrated)-248.4(that)-242(E134Bf)]TJ
T*
-.0201 Tc
[(e)-5.1(x)1.8(erts)-280.2(A)-4.8(D)1(CC)-281.4(and)-280.4(C).8(DC)-281.4(a)-5.1(g)1.8(ainst)-280.5(d)-3.9(EGFR-e)-5.1(xpressing)-280.4(SNP)-280.2(c)-5.1(ells.)]TJ
/F9 1 Tf
8.9663 0 0 8.9663 310.8472 259.37 Tm
0 Tc
[(Antitumor)-335.1(activities)-334.2(of)-336.7(E134Bf)-328.8(in)-336.5(the)-334.8(mouse)]TJ
0 -1.2203 TD
[(xenografts)-333.8(of)-330.4(SNP)-333.7(cells)]TJ
/F4 1 Tf
9.843 0 0 9.843 320.8251 231.9873 Tm
[(In)-198(the)-206.2(SNP)-199.7(xenograft)-202.4(models,)-208.2(E134Bf)-198.9(\(100)]TJ
/F10 1 Tf
17.1985 0 TD
(l)Tj
/F4 1 Tf
.5472 0 TD
[(g\))-203.8(and)-200.3(control)]TJ
-18.7594 -1.1116 TD
[(dog)-193.4(IgG)-194.2(\(100)]TJ
/F10 1 Tf
5.4372 0 TD
(l)Tj
/F4 1 Tf
.5529 0 TD
[(g\))-192.2(were)-197.6(injected)-197(intraperitoneally)-202.6(into)-194.7(mice)-198.2(on)]TJ
-5.9901 -1.1174 TD
[(days)-287.6(6,)-286.8(15,)-287.9(and)-292.4(23)-284.4(following)-293.4(the)-286.8(injection)-289.2(of)-290.2(SNP)-286.1(cells.)-293.6(Ca-)]TJ
0 -1.1116 TD
[(nine)-236.1(MNC)-242.3(were)-238(also)-235.9(injected)-243.1(surrounding)-242.4(the)-235(tumors)-238.1(in)-238.5(both)]TJ
T*
[(groups)-317.4(on)-319(days)-322.2(6,)-321.3(15,)-316.7(and)-321.2(23.)-316.7(The)-322.4(tumor)-317.9(volume)-320.2(was)-321.9(mea-)]TJ
0 -1.1174 TD
[(sured)-231.1(on)-232.6(days)-230(6,)-229.2(12,)-230.3(15,)-230.3(17,)-236(23,)-230.3(26,)-230.3(and)-229.1(28)-232.6(after)-232.5(the)-229.2(injection.)]TJ
0 -1.1116 TD
[(The)-408.8(administration)-416.8(of)-411.1(E134Bf)-412(resulted)-414.3(in)-405.5(a)-411.2(signicant)-416.6(re-)]TJ
T*
[(duction)-314.4(in)-307.6(tumor)-312.1(development)-316.6(on)-313.2(day)-309.7(12)-313.2(\()]TJ
/F11 1 Tf
17.498 0 TD
(p)Tj
/F8 1 Tf
.6681 0 TD
(<)Tj
/F4 1 Tf
.72 0 TD
[(0.05\),)-310.7(day)-315.5(15)]TJ
-18.8861 -1.1174 TD
(\()Tj
/F11 1 Tf
.5011 0 TD
(p)Tj
/F8 1 Tf
.6624 0 TD
(<)Tj
/F4 1 Tf
.72 0 TD
[(0.01\),)-535.4(day)-540.1(17)-537.8(\()]TJ
/F11 1 Tf
6.8886 0 TD
(p)Tj
/F8 1 Tf
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(<)Tj
/F4 1 Tf
.72 0 TD
[(0.01\),)-535.4(day)-540.1(23)-532.1(\()]TJ
/F11 1 Tf
6.8886 0 TD
(p)Tj
/F8 1 Tf
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(<)Tj
/F4 1 Tf
.72 0 TD
[(0.01\),)-535.4(day)-540.1(26)]TJ
-18.4368 -1.1116 TD
(\()Tj
/F11 1 Tf
.5011 0 TD
(p)Tj
/F8 1 Tf
.6624 0 TD
(<)Tj
/F4 1 Tf
.72 0 TD
[(0.01\),)-322.3(and)-321.3(day)-327(28)-319(\()]TJ
/F11 1 Tf
8.0175 0 TD
(p)Tj
/F8 1 Tf
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(<)Tj
/F4 1 Tf
.7142 0 TD
[(0.01\))-324.6(compared)-324.5(with)-322.3(that)-322.6(of)-324.7(the)]TJ
-11.2833 -1.1116 TD
[(control)-279.8(dog)-279.8(IgG)-280.6(\(Fig.)-284.2(3\).)]TJ
1.0137 -1.1174 TD
[(The)-241.8(administration)-244(of)-244.1(E134Bf)-245(resulted)-241.6(in)-244.2(a)-244.2(35%)-240.5(reduction)]TJ
-1.0137 -1.1116 TD
[(of)-284.4(tumor)-283.3(volume)-285.6(compared)-290(with)-282(that)-288.1(of)-284.4(the)-286.8(control)-285.6(dog)-285.5(IgG)]TJ
T*
[(on)-192.3(day)-188.8(28)-192.3(postinjection.)-199.1(Furthermore,)-197.5(the)-188.9(tumor)-196.9(weight)-193.2(of)-192.2(the)]TJ
T*
[(E134Bf-treated)-476.3(mice)-468.9(was)-471.7(signicantly)-470.7(lower)-468.4(than)-472.2(that)-472.4(of)]TJ
0 -1.1174 TD
[(control)-625.4(dog)-619.6(IgG-treated)-632.9(mice)-624.4(\(28%)-621.7(reduction;)]TJ
/F11 1 Tf
20.902 0 TD
(p)Tj
/F8 1 Tf
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(<)Tj
/F4 1 Tf
.72 0 TD
(0.05,)Tj
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[(Fig.)-248.6(4A\).)-249.5(The)-253.3(resected)-255.3(tumors)-249.7(are)-253.3(shown)-253.9(in)-250(Figure)-249.6(4B.)-248.6(Total)]TJ
ET
q
1 i
328.252 732.245 m
532.573 732.245 l
532.573 386.872 l
328.252 386.872 l
W n
q
204.461 0 0 345.373 328.252 386.872 cm
/Im2 Do
Q
Q
BT
/F5 1 Tf
9.845 0 0 9.845 310.8472 370.0912 Tm
[(FIG.)-544(1.)]TJ
/F4 1 Tf
4.3247 0 TD
[(Flow)-540.3(cytometry)-548.3(using)-540.5(E134Bf.)]TJ
/F5 1 Tf
13.8263 0 TD
(\(A\))Tj
/F4 1 Tf
1.9234 0 TD
[(SNP)-544.7(cells)]TJ
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[(were)-473.6(treated)-471.5(with)-471.6(E134Bf)-474.5(or)-468.4(buffer)-473.6(control,)-472.5(followed)-476.7(by)]TJ
T*
[(FITC-conjugated)-510.2(anti-dog)-504.7(IgG.)]TJ
/F5 1 Tf
13.5671 0 TD
(\(B\))Tj
/F4 1 Tf
1.8427 0 TD
[(Determination)-508.5(of)-508.7(the)]TJ
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[(binding)-479.5(afnity)-480.5(of)-485.7(E134Bf)-480.3(for)-480.9(SNP)-481.4(cells)-485.4(using)-482.9(ow)-481.6(cy-)]TJ
0 -1.0077 TD
[(tometry.)-467.9(SNP)-464.1(cells)-468.2(were)-467.8(suspended)-471.6(in)-462.8(100)]TJ
/F12 1 Tf
18.3985 0 TD
(m)Tj
/F4 1 Tf
.5471 0 TD
[(L)-468.6(o)0(f)-468.4(serially)]TJ
-18.9456 -1.0135 TD
[(diluted)-814.8(E134Bf,)-817.7(followed)-822.2(by)-813.9(the)-816.3(addition)-821.6(of)-813.9(FITC-)]TJ
T*
[(conjugated)-365(anti-dog)-366.5(IgG.)-364.2(Fluorescence)-368.8(data)-369.3(were)-364.2(collected)]TJ
0 -1.0077 TD
[(using)-333.2(the)-332.6(EC800)-335.3(Cell)-334.7(Analyzer.)]TJ
/F6 1 Tf
8.9663 0 0 8.9663 59.7543 749.7069 Tm
[(DEFUCOSYLATED)-341.8(MOUSE-DOG)-336.8(CHIMERIC)-334(ANTI-EGFR)]TJ
6.4556 0 0 6.4556 300.8125 749.7069 Tm
(M)Tj
8.9663 0 0 8.9663 306.1983 749.7069 Tm
(A)Tj
6.4556 0 0 6.4556 312.6046 749.7069 Tm
(B)Tj
8.9663 0 0 8.9663 540.0565 749.7069 Tm
(55)Tj
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