CN117069828A

CN117069828A - Preparation and application of anti-herpes zoster virus gE protein monoclonal antibody

Info

Publication number: CN117069828A
Application number: CN202310803976.1A
Authority: CN
Inventors: 张翊; 罗煊; 王顺强; 周立娟; 姚健; 简建升; 闫妙妙; 朱晨; 井申荣
Original assignee: Zhejiang Tianyuan Biological Pharmaceutical Co ltd
Current assignee: Zhejiang Tianyuan Biological Pharmaceutical Co ltd
Priority date: 2023-07-03
Filing date: 2023-07-03
Publication date: 2023-11-17

Abstract

The invention provides an anti-herpes zoster virus gE protein monoclonal antibody, which comprises an antibody 1, wherein the antibody 1 comprises a light chain and a heavy chain; the light chain comprises a light chain variable region and a light chain constant region; the light chain variable region includes the following three complementarity determining region CDRs: SEQ ID NO:1, CDR1, SEQ ID NO:2, CDR2, SEQ ID NO:3, CDR3; the light chain variable region has the sequence shown in SEQ ID NO:13, an amino acid sequence shown in seq id no; the heavy chain comprises a heavy chain variable region and a heavy chain constant region; the heavy chain variable region includes the following three complementarity determining region CDRs: SEQ ID NO:4, CDR1, SEQ ID NO:5, CDR2, SEQ ID NO: CDR3 shown in fig. 6; the heavy chain variable region has the amino acid sequence of SEQ ID NO:14, and a polypeptide having the amino acid sequence shown in seq id no.

Description

Preparation and application of anti-herpes zoster virus gE protein monoclonal antibody

Technical Field

The invention relates to the field of medicines, in particular to preparation and application of an anti-herpes zoster virus gE protein monoclonal antibody.

Technical Field

Varicella-zoster virus (varicella zoster virus, VZV) belongs to alpha-type herpesvirus, is a double-stranded DNA virus with a capsule structure, and mainly has glycoprotein, envelope, nucleocapsid and DNA, and the diameter is only between 180 and 200 nm.

The primary infection of the VZV has a typical winter and spring seasonal occurrence mode, the infectivity is extremely strong, varicella is caused by primary infection virus, the VZV is hidden in nerves during the period, and when immunity is reduced due to the reasons of aging, emotional stress, drug treatment and the like, the VZV is activated again, so as to cause herpes zoster.

Shingles (HZ) caused by VZV infection is an acute infectious skin disease, primary infected population is mainly infants under 6 years old, varicella is caused after infection, varicella belongs to self-limiting disease, but VZV virus is hidden in nerve cells, and when individual immunity is low, VZV virus is replicated and proliferated again, thereby causing shingles, inflammation and cell death in tissues subject to related neurons, which is accompanied with persistent nerve root Pain (PHN) for a short month and several decades, so it is called as dead cancer, and the specific occurrence mechanism of postherpetic neuralgia is unknown and treatment is difficult.

Shingles are widely prevalent worldwide, with prevalence increasing year by year and a trend toward younger age, and also show a strong seasonal pattern, with prevalence mainly concentrated in spring and winter, with 1.4 million new cases per year worldwide statistically, 420 tens of thousands leading to serious complications of hospitalization and 4200 deaths. In addition, the age of patients is mostly focused on ages 50 and older, and women have a larger proportion than men, so age is considered to be the most important risk factor for HZ infection. Although mortality after illness is low, patients often suffer from tremendous physical pain, and surveys show that at least 50% of the elderly have or are suffering from shingles, with PHN morbidity up to 80% in people over 50 years old, with severe physical and psychological distress and loss of property for the patient and family members.

The VZV gE, gB and gH can induce the organism to generate neutralizing antibodies, the gE has the largest molecular weight and neutralizing related epitopes, and the highest content on the viral envelope is the main viral antigen and is also the main candidate antigen for preparing the subunit viral vaccine. At present, no detection kit and no report of a monoclonal antibody sequence thereof are disclosed for detecting the gE protein, and no genetic engineering technology is adopted to prepare the gE protein as a vaccine and information for preventing herpes zoster virus infection and herpes zoster diseases is marketed, so that the invention discloses the preparation of the monoclonal antibody for resisting the gE protein and the sequence thereof, and has important significance for developing a more effective method for early diagnosis of VZV virus and a kit for qualitatively and quantitatively measuring the gE protein content.

Disclosure of Invention

The invention aims to provide a preparation method and application of an anti-herpes zoster virus gE protein monoclonal antibody.

A polynucleotide 1 encoding a polypeptide comprising: the light chain variable region, light chain, heavy chain variable region, heavy chain of claim 1.

A vector 1 comprising the polynucleotide 1 of claim 2.

A genetically engineered cell 1 comprising the polynucleotide 1 of claim 2 or the vector 1 of claim 3.

An anti-herpes zoster virus gE protein monoclonal antibody comprising an antibody 2, said antibody 2 comprising a light chain and a heavy chain; the light chain comprises a light chain variable region and a light chain constant region, and the heavy chain comprises a heavy chain variable region and a heavy chain constant region; the light chain variable region includes the following three complementarity determining region CDRs: SEQ ID NO:7, CDR1, SEQ ID NO:8, CDR2, SEQ ID NO: CDR3 as shown in 9; the light chain variable region has the sequence shown in SEQ ID NO:15, and a polypeptide comprising the amino acid sequence shown in seq id no; the heavy chain variable region includes the following three complementarity determining region CDRs: SEQ ID NO:10, CDR1, SEQ ID NO:11, CDR2, SEQ ID NO:12, CDR3; the heavy chain variable region has the amino acid sequence of SEQ ID NO:16, and a polypeptide having the amino acid sequence shown in seq id no.

A polynucleotide 2 encoding a polypeptide comprising: the light chain variable region, light chain, heavy chain variable region, heavy chain of claim 5.

A vector 2 comprising the polynucleotide 2 of claim 6.

A genetically engineered cell 2 comprising the polynucleotide 2 of claim 6 or the vector 2 of claim 7.

A kit, comprising: the antibody 1 of claim 1, and/or the antibody 2 of claim 5.

An immunoconjugate comprising an antibody and a conjugate; the antibody comprises the antibody 1 of claim 1, or the antibody 2 of claim 5; the conjugate includes one or more of a fluorescein, a drug, a toxin, a cytokine, a radionuclide, or an enzyme.

In order to obtain the anti-herpes zoster virus gE protein monoclonal antibody, the gE protein prepared by the company is mixed with an adjuvant, mice are immunized for multiple times, high-titer polyclonal antibodies and mouse spleens are obtained, animal spleens are taken out, spleen cells are separated, in vitro fusion with NS0 tumor cells is carried out, hybridoma cell strains of the monoclonal antibodies are screened, antibody sequences of the anti-gE protein are obtained for the hybridoma cells with high secretion of the monoclonal antibodies through RT-PCR, the monoclonal antibody sequences are recombined onto pcDNA3.1 plasmid, CHO cells are transfected, the antibodies are expressed, and the obtained monoclonal antibody gene sequences can be combined with the gE protein through verification.

The related technology of the invention has the following beneficial effects:

1. the pair of monoclonal antibodies obtained by screening can recognize denatured and non-denatured VZV gE proteins, and can be used in immunoblotting detection and ELISA experiments.

2. The monoclonal antibody and the gene sequence thereof obtained by the invention can be prepared by adopting various known technologies (such as hybridoma technology and antibody engineering technology) by a person skilled in the art, and can also be prepared into a Fab segment by adopting a small antibody technology.

3. The pair of monoclonal antibodies of the invention are high-specificity and high-sensitivity raw materials, and can be directly used for detecting human herpes zoster virus or gE protein antigens, thereby being beneficial to early screening of virus infection, qualitative and quantitative detection of antigens.

Drawings

FIG. 1 is hybridoma cell growth following cell fusion;

FIG. 2 is a clone of the light and heavy chain genes of an anti-gE antibody, lane 1 is DNA Marker DL2000; lane 2 is GAPDH; lane 3 is light chain LC; lane 4 is heavy chain HC;

FIG. 3 is a cut-out of recombinant vector, lane 1 is a cut-out of LC-pMD-18T (large fragment of approximately 2711bp, small fragment 1380 bp); lane 2 is the restriction enzyme identification of HC-pMD-18T (approximately 2711bp for the large fragment, 780bp for the small fragment); m is DNA Marker DL5000;

FIG. 4 is a standard curve for detecting gE protein concentration for mab 1 and HRP-mab 2.

Detailed Description

The following examples are illustrative of the invention and are not intended to limit the scope of the invention. Unless otherwise indicated, the technical means employed in the examples are conventional means well known to those skilled in the art, and all raw materials used are commercially available.

EXAMPLE 1 gE protein immunized animals and hybridoma cell screening to secrete anti-gE specific monoclonal antibodies

Recombinant herpes zoster virus gE protein (Zhejiang Tianyuan Biopharmaceutical Co., ltd.) was mixed with Freund's complete adjuvant (purchased from Sigma Co.), and BALB/c mice (purchased from Venlhua laboratory animal technologies Co., ltd.) were vaccinated four times at 15-day intervals. One week after the last booster immunization, mice were bled from the tail vein and ELISA was performed to determine antibody titers.

The day before hybridoma cells were prepared, the non-immunized BALB/c mice were collected by eyeball blood as negative control serum, and then, after being killed by removal of the neck, they were placed in a beaker containing 75% alcohol and immersed for 5 minutes, the mice were placed in a 10cm sterile plate, approximately 5mL of prepared HAT culture solution (purchased from Sigma Co.) was slowly injected into the abdominal cavity using a 10mL sterile syringe, the bellies of the mice were gently rubbed, and the syringe was used to aspirate the culture solution containing macrophages in the abdominal cavity and collected in a sterile 50mL centrifuge tube, and repeated 2 times. Finally, HAT culture solution was added to the centrifuge tube to 50mL, gently pipetted several times, plated at 100. Mu.L/well (96 well cell culture plate), and placed in a carbon dioxide incubator for culturing.

Resuscitates myeloma cell NS0 (purchased from Shenzhen Shengzhen biological Co., ltd.) to ensure that the myeloma cell activity rate is over ninety percent during fusion, takes out myeloma cells 1000rmp in the logarithmic phase of growth, centrifugates for 10min, removes the culture medium supernatant, resuspents cells with serum-free culture medium, counts and spare.

Mice after the immunization were subjected to eyeball blood collection, and positive control serum was left. Mice were sacrificed by cervical removal and immersed in 75% alcohol for 5min. Mice were placed in 10cm sterile dishes, spleens of the mice were peeled off with sterile scissors and forceps, placed on a cell strainer, sheared and thoroughly ground, and spleen cells were collected. Centrifugation at 1000rmp for 10min, discarding supernatant, re-suspending cells with serum-free medium, centrifuging, and cell counting for use.

Spleen cells and myeloma cells are uniformly mixed according to the ratio of 9:1, culture solution is added to 50mL,1000rmp is centrifuged for 10min, the supernatant is discarded, the bottom of the tube is tapped, and cell aggregates accumulated at the bottom are loosened. The centrifuge tube was placed in a beaker containing sterile water at 37 ℃. (1) 1mL of PEG (from Sigma Co.) was slowly added over 1min (pre-warmed at 37 ℃ C.) and the tube gently shaken during the addition, and allowed to stand for 1min after the addition. (2) The termination was performed by slowly adding 1mL of RPMI-1640 medium (purchased from Sigma Co.) over 1min. (3) 15mL of RPMI-1640 medium was slowly added over 15 min. (4) 1000rmp was centrifuged for 10min, the supernatant was discarded, 50mL of HAT medium was gently pipetted and mixed, 100. Mu.L/well was added to a 96-well plate containing feeder cells, and incubated in a 5% CO2 incubator at 37 ℃.

After 5 days following cell fusion, a small amount of cell clumping was seen in the 96-well plate (FIG. 1), and the 96-well plate was half-exchanged on day 7, 100. Mu.L of HAT medium was aspirated from the plate, and 100. Mu.L of HT medium (purchased from Sigma Co.) was added. After 10 days of fusion, the cell fusion results were observed and the cell supernatants of the well plates were subjected to indirect ELISA detection. Cell counting is carried out on cell holes positive to the antibody, then subcloning is carried out according to 0.5 cells/hole, and 20 hybridoma cells capable of secreting monoclonal antibody are obtained after three subcloning, and the hybridoma cells are subjected to expansion culture and frozen storage.

Example 2 typing and antibody pair analysis of anti-gE specific monoclonal antibodies

For the culture medium of 10 hybridoma cells secreting a relatively high amount of antibody after three rounds of subcloning screening, ELISA typing was performed using SBA mouse monoclonal antibody typing kit (purchased from southern Biotech Co., U.S.A., 5300-01) and the results are shown in Table 1. The typing results show that: the heavy chain types are mainly IgM, igG1, igG2a, and the light chain has only one kappa chain.

Sequence number	Antibody numbering	Heavy chain type	Light chain type
				1	2D1	IgM	kappa
2	2G5	IgG1	kappa
				3	3E10	IgM	kappa
4	4F9	IgG1	kappa
				5	7A3	IgG1	kappa
6	7C6	IgG1	kappa
				7	9D2	IgG1	kappa
8	9F8	IgG1	kappa
				9	11H10	IgG1	kappa
10	13G7	IgG2a	kappa

TABLE 1 subtype of anti-gE monoclonal antibodies

And respectively pairing the 6 monoclonal antibodies pairwise, and judging whether the monoclonal antibodies recognize the same or similar epitope according to ELISA (enzyme-linked immunosorbent assay) addition experiments. ELISA experiments were performed to determine OD450 values by coating gE antigen, adding two monoclonal antibodies as primary antibodies in one well, and HRP-labeled goat anti-mouse IgG antibodies (purchased from Abclonal corporation) as secondary antibodies. The addition index AI was calculated by the formula AI (%) = [2×a (1+2)/(a1+a2) -1] ×100%, where A1, A2 and a (1+2) represent the OD450 values measured for the 1 st, 2 nd and two monoclonal antibody mixtures, respectively, when AI is not less than 50%, it was confirmed that there was no competition relationship between the two antibodies, and when AI was less than 50%. The experimental results are shown in table 2, wherein the AI value is the largest when the 7C6 antibody and the 9F8 antibody are paired, and it is proved that there is no competition between the two antibodies, and the antibodies can be used as an ELISA kit for constructing the antibody pair.

Antibodies to

2D1

2G5

4F9

7C6

9F8

11H10

2D1

－

67.5

42.7

89.3

50.2

35.4

2G5

－

80.4

44.1

23.1

62.5

4F9

－

27.5

82.6

42.9

7C6

－

92.3

38.6

9F8

－

75.3

11H10

－

TABLE 2.6 pairing analysis of monoclonal antibodies

EXAMPLE 3.2 Gene cloning and sequencing analysis of monoclonal antibodies

Screening, wherein 2 strains of 7C6 and 9F8 monoclonal antibodies can be used for quantitatively detecting ELISA antibody pairs of gE protein content, performing amplification culture on hybridoma cells secreting and expressing 7C6 and 9F8 monoclonal antibodies, centrifuging at a low speed, discarding culture solution, suspending by using PBS, adding Trizol reagent (purchased from Novoin company) solution, mixing uniformly, adding chloroform, shaking, standing at room temperature for 10min, centrifuging at a low temperature of 12000rpm for 10min, sucking supernatant water into the other 1.5ml, adding NaAc and isopropanol, mixing uniformly for 10min at-70 ℃, centrifuging at a low temperature of 12000rpm for 10min after taking out, discarding supernatant of a centrifuge tube, suspending precipitated RNA by using 10 μl sterile purified water, and preserving at-70 ℃ for later use.

Using PrimeScript ^TM The One Step RT-PCR kit (purchased from Takara) was used to amplify the light and heavy chain gene fragments of the antibody, and the results were shown in FIG. 2. And the light and heavy chain genes of the antibody were ligated to pMD-18T vector (purchased from Takara corporation) according to the protocol. Identification of the correct recombinant vector (FIG. 3) was submitted to Shanghai Ind sequencing.

Comparing the determined variable region sequences of the two antibodies with the variable region gene sequences of the heavy chain and the light chain of the antibodies in a database IGBLAST (https:// www.ncbi.nlm.nih.gov/IGBLAST /), the heavy chain gene and the light chain gene of the antibodies are consistent with the framework sequence of the antibodies, the consistency ratio is more than 80 percent, and the antibody is the light chain gene and the heavy chain gene of the antibody. The amino acid sequence comparison of the antibody shows that the amino acid sequence of the light chain and the heavy chain of the antibody has the consistency of less than 40 percent with the amino acid sequence of the prior monoclonal antibody, which indicates that the monoclonal antibody prepared by the invention has not been reported yet.

The heavy and light chain sequences of the 7C6 and 9F8 mabs are as follows (single underlined is the variable region sequence, dashed underlined is the constant region sequence).

7C6 mab light chain amino acid sequence:

DIVLTQSPLSLPVSLGDQASISCRSSQSIIHSNGDTYLEWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHVPWTFGGGTKLEIKRADAAPTVSIFPPSSEQSTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC

7C6 monoclonal antibody heavy chain amino acid sequence:

QVQLEQSGGGLVKPGGSLKLSCAASGFTFSDYYIYWVRQTPEKRLEWVATISDGGSYTYYPDSVKGRFTISRDNTKNNLYLQMSSLKSEDTAMYYCTRAGGQAAWFAYWGQGTLVTVSAAKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK

9F8 mab light chain amino acid sequence:

DIVMTQSPASLAVSLGQRATISYRASKSVSTSGYSYMHWNQQKPGQPPRLLIYLVSNLESGVPARFSGSGSGTDFTLNIHPVEEEDAATYYCQHIRELTRSEGGPSWKQRADAAPTLSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC

9F8 monoclonal antibody heavy chain amino acid sequence:

EVKLEQSGPELVKPGASMKMSCKASGYTFTSYVIHWVKQKPGQGLEWIGYIDPYNDGTKYNEKFRGKATLTSDKSSSTAYMELSSLTSEDSAVFYCARGGYYAMDYWGQGTSVTVSSAKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPESVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVSFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQV

YTIPPPKKQMAKDKVSLTCMITDFFPEDITVEWQWNGQPAENYKNTQPIMDTDGSYF

VYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK

the CDR-region amino acid sequences and nucleotide sequences of the variable regions of monoclonal antibody 1 (7C 6) and monoclonal antibody 2 (9F 8) are summarized in table 3.

TABLE 3 summary of CDR sequences of monoclonal antibodies

EXAMPLE 4.2 expression and purification of monoclonal antibodies

To verify that the cloned genes for the 7C6 and 9F8 monoclonal antibodies were correct, the heavy and light chain coding sequences were recombined into pcDNA6 (available from ThermoFish Co.) respectively, and expression vectors pcDNA6-H (7C 6) and pcDNA6-L (7C 6), pcDNA6-H (9F 8) and pcDNA6-L (9F 8) were constructed, and the methods were performed as described herein using the ExpiFectamine transfection reagent to co-transfect ExpiCHO-S cells (available from ThermoFish Co.). After 3 days, the cells were transferred to 300ml of an ExpiCHO expression medium (available from ThermoFish corporation), shake flask culture was performed at 37℃in a shaker at 120rpm, and supernatants were harvested after 6, 8, 10, 12 days of feeding, and 14 days of culture. Cell supernatants (diluted 1000-fold) were assayed by ELISA for the presence of antibodies that specifically bind to gE protein. Table 4 shows that the cell supernatant expressing the 2 monoclonal antibody sequences has very high absorbance; the fact that none of the control cell supernatants without the transfection of the relevant plasmid had a binding signal indicates that the 7C6 and 9F8 mab gene sequences prepared according to the invention still have biological activity for binding to gE protein after expression in animal cells.

Sample of	ELISA OD450 value
		Positive control	2.608±0.162
Negative control	0.165±0.051
		7C6-ExpiCHO-S cell supernatant	2.125±0.181
9F8-ExpiCHO-S cell supernatant	1.832±0.206
		ExpiCHO-S cell supernatant	0.133±0.069

TABLE 4 determination of expression of monoclonal antibodies by transfected CHO cells

EXAMPLE 5 establishment and application of ELISA Sandwich method kit

And preparing and purifying ascites, namely taking 8-10-week-old female BALB/c mice, performing intraperitoneal injection of 500 mu L of Freund's incomplete adjuvant on each mouse for sensitization for 5-7 days, and then performing intraperitoneal injection of hybridoma cells with good growth state and over 90% of activity rate. After 5-10 days, the abdomen of the mice is obviously swelled, and ascites is collected. The ascites is centrifuged for 15min at a low temperature of 4500rpm, the supernatant is sucked, and monoclonal antibody is purified directly by Protein A affinity chromatography, and is packaged and frozen for standby after collection.

And (3) establishing an ELISA sandwich method kit, wherein the purified 7C6 monoclonal antibody is used as a coating antibody, and the 9F8 monoclonal antibody is used as a detection antibody after being marked by horseradish peroxidase. Coating the 7C6 monoclonal antibody with carbonate at the temperature of 96-well plates for overnight at 4 ℃, washing three times by PBST in the next day, sealing by using 5% skimmed milk powder-PBST, adding gE protein and detection antibody with different concentrations, washing three times by PBST at the temperature of 4 ℃ in the third day, adding a chromogenic liquid for light-proof color development, and detecting an OD value by an enzyme-labeled instrument at 450 nm. OD450nm is taken as ordinate and gE protein concentration is taken as abscissa, and a standard curve is drawn and four-parameter fitting analysis is performed. As shown in fig. 4, after four-parameter Logistic curve fitting, the equation of y= (a-D)/[ 1+ (x/C)/(B ] +d, where a=3.80223, b= -1.68271, c=0.40471, d=0.02715, r++2= 0.99941, indicates that the pair of monoclonal antibodies can be used in combination for qualitative and quantitative detection of gE protein.

Finally, it should also be noted that the above list is only one specific embodiment of the present invention. Obviously, the invention is not limited to the above embodiments, but many variations are possible. All modifications directly derived or suggested to one skilled in the art from the present disclosure should be considered as being within the scope of the present invention.

Claims

1. An anti-herpes zoster virus gE protein monoclonal antibody comprising an antibody 1, said antibody 1 comprising a light chain and a heavy chain;

the light chain comprises a light chain variable region and a light chain constant region;

the light chain variable region includes the following three complementarity determining region CDRs:

SEQ ID NO:1, CDR1, SEQ ID NO:2, CDR2, SEQ ID NO:3, CDR3;

the light chain variable region has the sequence shown in SEQ ID NO:13, an amino acid sequence shown in seq id no;

the heavy chain comprises a heavy chain variable region and a heavy chain constant region;

the heavy chain variable region includes the following three complementarity determining region CDRs:

SEQ ID NO:4, CDR1, SEQ ID NO:5, CDR2, SEQ ID NO: CDR3 shown in fig. 6;

the heavy chain variable region has the amino acid sequence of SEQ ID NO:14, and a polypeptide having the amino acid sequence shown in seq id no.

2. A polynucleotide 1, wherein said polynucleotide encodes a polypeptide that:

the light chain variable region, light chain, heavy chain variable region, heavy chain of claim 1.

3. A vector 1 comprising the polynucleotide 1 of claim 2.

4. A genetically engineered cell 1 comprising the polynucleotide 1 of claim 2 or the vector 1 of claim 3.

5. An anti-herpes zoster virus gE protein monoclonal antibody comprising an antibody 2, said antibody 2 comprising a light chain and a heavy chain;

the light chain comprises a light chain variable region and a light chain constant region, and the heavy chain comprises a heavy chain variable region and a heavy chain constant region;

SEQ ID NO:7, CDR1, SEQ ID NO:8, CDR2, SEQ ID NO: CDR3 as shown in 9;

the light chain variable region has the sequence shown in SEQ ID NO:15, and a polypeptide comprising the amino acid sequence shown in seq id no;

SEQ ID NO:10, CDR1, SEQ ID NO:11, CDR2, SEQ ID NO:12, CDR3;

the heavy chain variable region has the amino acid sequence of SEQ ID NO:16, and a polypeptide having the amino acid sequence shown in seq id no.

6. A polynucleotide 2, wherein said polynucleotide encodes a polypeptide that:

the light chain variable region, light chain, heavy chain variable region, heavy chain of claim 5.

7. A vector 2 comprising the polynucleotide 2 of claim 6.

8. A genetically engineered cell 2 comprising the polynucleotide 2 of claim 6 or the vector 2 of claim 7.

9. A kit, comprising: the antibody 1 of claim 1, and/or the antibody 2 of claim 5.

10. An immunoconjugate, characterized in that the immunoconjugate comprises an antibody and a conjugate;

the antibody comprises the antibody 1 of claim 1, or the antibody 2 of claim 5;

the conjugate includes one or more of a fluorescein, a drug, a toxin, a cytokine, a radionuclide, or an enzyme.